Challenges and opportunities in neurometabolic disease treatment with enzyme delivery.

INTRODUCTION: Neurometabolic disorders remain challenging to treat, largely due to the limited availability of drugs that can cross the blood-brain barrier (BBB) and effectively target brain impairment. Key reasons for inadequate treatment include a lack of coordinated knowledge, few studies on BBB status in these diseases, and poorly designed therapies. AREAS COVERED: This paper provides an overview of current research on neurometabolic disorders and therapeutic options, focusing on the treatment of neurological involvement. It highlights the limitations of existing therapies, describes innovative protocols recently developed, and explores new opportunities for therapy design and testing, some of which are already under investigation. The goal is to guide researchers toward innovative and potentially more effective treatments. EXPERT OPINION: Advancing research on neurometabolic diseases is crucial for designing effective treatment strategies. The field suffers from a lack of collaboration, and a strong collective effort is needed to enhance synergy, increase knowledge, and develop a new therapeutic paradigm for neurometabolic disorders.

Opportunities for nanomaterials in enzyme therapy.

In recent years, enzyme therapy strategies have rapidly evolved to catalyze essential biochemical reactions with therapeutic potential. These approaches hold particular promise in addressing rare genetic disorders, cancer treatment, neurodegenerative conditions, wound healing, inflammation management, and infectious disease control, among others. There are several primary reasons for the utilization of enzymes as therapeutics: their substrate specificity, their biological compatibility, and their ability to generate a high number of product molecules per enzyme unit. These features have encouraged their application in enzyme replacement therapy where the enzyme serves as the therapeutic agent to rectify abnormal metabolic and physiological processes, enzyme prodrug therapy where the enzyme initiates a clinical effect by activating prodrugs, and enzyme dynamic or starving therapy where the enzyme acts upon host substrate molecules. Currently, there are >20 commercialized products based on therapeutic enzymes, but approval rates are considerably lower than other biologicals. This has stimulated nanobiotechnology in the last years to develop nanoparticle-based solutions that integrate therapeutic enzymes. This approach aims to enhance stability, prevent rapid clearance, reduce immunogenicity, and even enable spatio-temporal activation of the therapeutic catalyst. This comprehensive review delves into emerging trends in the application of therapeutic enzymes, with a particular emphasis on the synergistic opportunities presented by incorporating enzymes into nanomaterials. Such integration holds the promise of enhancing existing therapies or even paving the way for innovative nanotherapeutic approaches.

Tumor-derived biomimetic nanozyme with immune evasion ability for synergistically enhanced low dose radiotherapy.

High doses of radiation can cause serious side effects and efficient radiosensitizers are urgently needed. To overcome this problem, we developed a biomimetic nanozyme system (CF) by coating pyrite (FeS2) into tumor-derived exosomes for enhanced low-dose radiotherapy (RT). CF system give FeS2 with immune escape and homologous targeting abilities. After administration, CF with both glutathione oxidase (GSH-OXD) and peroxidase (POD) activities can significantly lower the content of GSH in tumor tissues and catalyze intracellular hydrogen peroxide (H2O2) to produce a large amount of ·OH for intracellular redox homeostasis disruption and mitochondria destruction, thus reducing RT resistance. Experiments in vivo and in vitro showed that combining CF with RT (2 Gy) can provide a substantial suppression of tumor proliferation. This is the first attempt to use exosomes bionic FeS2 nanozyme for realizing low-dose RT, which broaden the prospects of nanozymes.

Enhancing the therapeutic use of biofilm-dispersing enzymes with smart drug delivery systems.

Biofilm-dispersing enzymes degrade the extracellular polymeric matrix surrounding bacterial biofilms, disperse the microbial community and increase their susceptibility to antibiotics and immune cells. Challenges for the clinical translation of biofilm-dispersing enzymes involve their susceptibility to denaturation, degradation, and clearance upon administration in vivo. Drug delivery systems aim to overcome these limitations through encapsulation, stabilization and protection from the exterior environment, thereby maintaining the enzymatic activity. Smart drug delivery systems offer target specificity, releasing payloads at the site of infection while minimizing unnecessary systemic exposure. This review highlights critical advances of biofilm-dispersing enzymes as a novel therapeutic approach for biofilm-associated infections. We explore how smart, bio-responsive delivery systems overcome the limiting factors of biofilm-dispersing enzymes and summarize the key systems designed. This review will guide future developments, focusing on utilizing selective and specific therapies in a targeted fashion to meet the unmet therapeutic needs of biofilm infections.

Effect of different levels of bioplex manganese along with probiotics and multi-enzymes on performance and immune system indices of broilers.

To investigate the effect of different levels of bioplex manganese along with probiotics and multi-enzymes on the performance and immune system of broilers, 640 one-day-old male chicks of the Ross 308 strain were reared and the data analysed in a 4 × 2 × 2 factorial experiment with four levels of bioplex manganese (0, 60, 72 and 84 mg per kg of diet), two levels of Parsilact probiotic (0 and 200 mg per kg of diet) and two levels of Combo multi-enzyme (0 and 1,000 mg per kg of diet) in a completely randomized design with 16 experimental treatments, 4 replicates and 10 chickens per replicate during a period of 42 days. The results showed that the performance of the broiler chickens in the diets containing 72 and 84 mg bioplex manganese along with probiotics and multi-enzymes had the greatest difference compared to the control (p < .05). Compared to the control with 0 mg/kg manganese; the bursa of Fabricius weight was greater in chickens fed diets containing additional manganese (p < .05). The concentration of antibodies produced against Newcastle disease virus, as well as the concentrations of IgG, IgM and total immunoglobulins produced against SRBC, were highest in the group fed a diet containing 84 mg manganese along with probiotics and multi-enzymes (p < .05). The results show combining additional manganese with probiotics and multi-enzymes in chicken diets leads to better performance as well as a stronger immune system of chickens.

Virus-Derived Peptides for Hepatic Enzyme Delivery.

Recently, a lipopeptide derived from the hepatitis B virus (HBV) large surface protein has been developed as an HBV entry inhibitor. This lipopeptide, called MyrcludexB (MyrB), selectively binds to the sodium taurocholate cotransporting polypeptide (NTCP) on the basolateral membrane of hepatocytes. Here, the feasibility of coupling therapeutic enzymes to MyrB was investigated for the development of enzyme delivery strategies. Hepatotropic targeting shall enable enzyme prodrug therapies and detoxification procedures. Here, horseradish peroxidase (HRP) was conjugated to MyrB via maleimide chemistry, and coupling was validated by SDS-PAGE and reversed-phase HPLC. The specificity of the target recognition of HRP-MyrB could be shown in an NTCP-overexpressing liver parenchymal cell line, as demonstrated by competitive inhibition with an excess of free MyrB and displayed a strong linear dependency on the applied HRP-MyrB concentration. In vivo studies in zebrafish embryos revealed a dominating interaction of HRP-MyrB with scavenger endothelial cells vs xenografted NTCP expressing mammalian cells. In mice, radiolabeled 125I-HRP-MyrBy, as well as the non-NTCP targeted control HRP-peptide-construct (125I-HRP-alaMyrBy) demonstrated a strong liver accumulation confirming the nonspecific interaction with scavenger cells. Still, MyrB conjugation to HRP resulted in an increased and NTCP-mediated hepatotropism, as revealed by competitive inhibition. In conclusion, the model enzyme HRP was successfully conjugated to MyrB to achieve NTCP-specific targeting in vitro with the potential for ex vivo diagnostic applications. In vivo, target specificity was reduced by non-NTCP-mediated interactions. Nonetheless, tissue distribution experiments in zebrafish embryos provide mechanistic insight into underlying scavenging processes indicating partial involvement of stabilin receptors.

Exogenous enzymes and probiotics alter digestion kinetics, volatile fatty acid content and microbial interactions in the gut of Nile tilapia.

Sustainable aquafeed production requires fishmeal replacement, leading to an increasing use of plant-derived ingredients. As a consequence, higher levels of antinutritional substances, such as non-starch polysaccharides and phytate, are present in aquafeeds, with negative effects on fish performance, nutrient digestibility and overall gut health. To alleviate these negative effects, providing exogenous digestive enzymes and/or probiotics can be an effective solution. In this study, we tested the effect of dietary supplementation of enzymes (phytase and xylanase) and probiotics (three strains of Bacillus amyloliquefaciens) on nutrient digestion kinetics and volatile fatty acid content along the gut, and the distal gut microbiome diversity in Nile tilapia. Chyme volatile fatty content was increased with probiotic supplementation in the proximal gut, while lactate content, measured for the first time in vivo in fish, decreased with enzymes along the gut. Enzyme supplementation enhanced crude protein, Ca and P digestibility in proximal and middle gut. Enzymes and probiotics supplementation enhanced microbial interactions as shown by network analysis, while increased the abundance of lactic acid bacteria and Bacillus species. Such results suggest that supplementation with exogenous enzymes and probiotics increases nutrient availability, while at the same time benefits gut health and contributes to a more stable microbiome environment.

Uso do gérmen de milho desengordurado, com e sem adição de um complexo enzimático, em dietas para cães / [Use of defatted corngerm with and without the addition of an enzyme complex in dog diets]

Objetivou-se avaliar o coeficiente de digestibilidade aparente (CDA) dos nutrientes, a palatabilidade das dietas e as características fecais de cães alimentados com uma dieta controle e uma dieta contendo 20% de gérmen desengordurado (GD), com e sem adição de complexo enzimático (amilase, xilanase, betaglucanase e mananase). Para o experimento de digestibidade e das características fecais, foram utilizados 12 cães adultos, distribuídos em delineamento em blocos ao acaso, em esquema fatorial 2 x 2 (dieta x enzima). O segundo experimento avaliou a palatabilidade, por meio da primeira escolha e da razão de ingestão (RI) da dieta DC vs. 20% de GD, utilizando-se 16 cães. O teste de palatabilidade contou com três dias consecutivos, totalizando 48 repetições. A dieta com inclusão de 20% de GD teve os menores valores de CDA da MS, da EB e da EM (P<0,05). A inclusão do complexo enzimático melhorou o CDA da MS, da EB e da EM (P<0,05). Não foram observadas diferenças nas características fecais (P>0,05). Em relação à palatabilidade, os cães preferiram a dieta 20% de GD, tanto na primeira escolha como na RI (P<0,05). A inclusão de enzimas às dietas melhora a digestibilidade dos nutrientes e da EM, sendo um aditivo com potencial uso na alimentação de cães.(AU)

The objective was to evaluate the apparent digestibility coefficient (ADC) of nutrients, diet palatability and fecal characteristics of dogs fed diets containing degreased germ (DG), and a control diet (DC) - both with and without the addition of enzyme complex (amylase, xylanase, betaglucanase and mananase). For the digestibility and fecal characteristics experiment 12 adult dogs were used, distributed in a randomized block design, in a 2 x 2 factorial scheme (diet x enzyme). The second experiment evaluated palatability using the first choice and ingestion ratio (IR) of DC diet vs. 20%gD, using 16 dogs. The palatability test had three consecutive days, totaling 48 repetitions. The diet with inclusion of 20% DG had the lowest ADC values of DM, GE and ME (P <0.05). Inclusion of the enzyme complex improved ADC of DM, GE and ME (P <0.05). No differences in fecal characteristics were observed (P >0.05). Regarding palatability, dogs preferred the 20% DG diet in both first choice and IR (P <0.05). Inclusion of enzymes in diets improves nutrient digestibility and ME, being an additive with potential use in dog food.(AU)

Effects of dietary supplementation of compound enzymes on performance, nutrient digestibility, serum antioxidant status, immunoglobulins, intestinal morphology and microbiota community in weaned pigs.

The objective of this study was to evaluate the effects of compound enzymes (CE) (containing per g 375 U amylase, 2500 U protease, 4000 U xylanase and 150 U ß-glucanase) on performance, nutrient digestibility, serum antioxidant status, immunoglobulins, intestinal morphology, volatile fatty acids contents and microbiota community in weaned pigs. Seventy-two pigs (Duroc × Landrace × Yorkshire, weaned at d 28) with an average body weight of 8.49 ± 0.87 kg were allotted into two treatments with six replicate pens per treatment (three barrows and three gilts per pen) according to sex and body weight in a randomised complete block design. The treatments contained a corn-soybean meal-barley basal diet (CON) or a basal diet supplemented with 1000 mg CE/kg (CE). The study was divided into phase 1 (d 1 to 14) and 2 (d 15 to 35). The average daily gain was increased (p < 0.05) in pigs fed CE in phase 2 and overall (d 1 to 35) compared with CON. These pigs had greater (p ≤ 0.05) serum IgA, IgG, superoxide dismutase and catalase contents, as well as tended to increase serum IgM content and apparent total tract digestibility (ATTD) of organic matter in phase 1 compared with CON. In phase 2, pigs supplemented with CE showed greater (p < 0.01) ATTD of dry matter, organic matter, crude protein and gross energy compared with CON. These pigs also had increased (p < 0.05) IgA, IgG, IgM, superoxide dismutase contents, and decreased (p < 0.05) malondialdehyde content in serum compared with CON. Moreover, pigs fed CE had higher (p < 0.05) villus height and villus height to crypt depth ratio in ileum, and tended to increased acetic acid content in colon compared with CON. Furthermore, pigs fed CE had increased (p < 0.05) relative abundance of Firmicutes at phylum level, Lactobacillales at order level, Lactobacillaceae at family level, Bacilli at class level, Lactobacillus at genus level in caecum and colon, as well as lower (p < 0.05) relative abundance of Bacteroidetes at phylum level, Bacteroidales at the order level, Bacteroidia at class level, Clostridium_sensu_stricto_6 at genus level in colon compared with CON. In conclusion, dietary inclusion of compound enzymes could effectively improve nutrient digestibility, serum antioxidant status, immunoglobulin, gut morphology, microbiota community, and therefore improve performance in weaned pigs.

Clinical potential of a rationally engineered enzyme for treatment of cocaine dependence: Long-lasting blocking of the psychostimulant, discriminative stimulus, and reinforcing effects of cocaine.

It is a grand challenge to develop a truly effective treatment of substance use disorder (SUD), particularly for cocaine and other drugs without an FDA-approved treatment available, because a truly effective therapy must effectively block the drug's physiological and reinforcing effects during the entire period of treatment in order to achieve the long-time abstinence required by the FDA. Whether a biologic, such as monoclonal antibody, vaccine, or therapeutic enzyme, can be truly effective for SUD treatment or not has been the subject of extensive debate. The main debate question is whether a biologic, particularly an exogenous enzyme, can effectively block the drug's reinforcing effect. In this report, we demonstrate that a modest dose of a recently redesigned long-acting cocaine hydrolase, CocH3-Fc(M6), can be used to effectively block the psychostimulant, discriminative stimulus, and reinforcing effects of cocaine for a sufficiently long period of time. For example, a dose of 3 mg/kg CocH3-Fc(M6) completely blocked the discriminative stimulus and reinforcing effects for 24/25 days and continued to significantly attenuate/decrease the cocaine effects for at least 29 days in rats. All the animal data consistently suggest that the long-acting cocaine hydrolase is a truly promising candidate of enzyme therapy for treatment of cocaine use disorder.

Características do trato digestivo, metabolizabilidade e retenção de nutrientes em frangos de corte alimentados com complexo enzimático / Characteristics of the digestive tract, metabolizability and nutrient retention in broilers fed enzymatic complex

The objective was to evaluate the digestive tract characteristics, metabolizability and nutrient retention of broilers fed diets supplemented with enzyme complex (EC). To evaluate the characteristics of the digestive tract 600 female Cobb 500 birds were used, distributed in a completely randomized design, with 5 inclusion levels of the EC (0; 100, 200, 300 and 400 g/ton) and 6 replicates of 20 birds each. To evaluate the metabolizability and the retention of nutrients 200 female Cobb 500 birds at 15 days of age were used, distributed in a completely randomized design with 5 levels of supplementation of the EC and 4 replicates of 10 birds each. No significant effects (P>0.05) were observed for the supplementation of the EC in the intestinal pH, digestive organ weight, intestinal length and metabolizable coefficients of dry matter and crude protein. The metabolizable coefficient of ethereal extract was influenced in a quadratic decreasing form (P<0.01). The metabolizable coefficients of calcium (Ca) and phosphorus (P) were influenced in a quadratic increase (P<0.01), resulting in increased Ca retention in 21.39% and P in 9.56%. Supplementation of the EC in broiler diets improves the metabolizability and retention of P and Ca, without affecting the other parameters evaluated.(AU)

Effects of feeding broiler breeder hens a coextruded full-fat flaxseed and pulses mixture without or with multienzyme supplement.

The effects of coextruded full-fat flaxseed and pulses (FFF; 1:1 wt/wt) mixture on n-3 polyunsaturated fatty acids (PUFA) enrichment in egg yolk, hepatic attributes, apparent retention (AR) of components, and ceca metabolites were evaluated in broiler breeder hens. The diets were as follows: 1) corn-soybean control, 2) control diet plus 18% FFF (FFF-), and 3) FFF plus enzyme supplement (FFF+) containing galactanase, protease, mannanase, glucanase, xylanase, amylase, and cellulase activities. Twenty-six-week-old Cobb 500 broiler breeder hens were allocated to 30 identical cages (2 hens/cage) and given 1-week adaptation period. The 3 diets were assigned to 10 replicate cages based on postadaptation BW and fed based on breeder curve for 30 D. Excreta samples were collected from day 24 to 27 for determination of AR of components, and eggs were collected from day 28 to 30 for yolk polyunsaturated fatty acids analyses. On day 30, birds were weighed, killed via cervical dislocation, liver weighed, and stored for fat analyses. Ceca digesta samples were taken for concentration of short-chain fatty acids. Liver and yolk weights as well as total yolk FA were not influenced by diets (P > 0.05). Control birds had lower yolk concentration of α-linolenic acid than birds fed either FFF- or FFF+ (P < 0.01) corresponding to 7.5, 36.8, and 37.3 mg/g for the control, FFF-, and FFF+, respectively. Control birds also exhibited lower yolk concentration of docosahexaenoic acid (P < 0.01). Control birds had higher hepatic concentration of crude fat and apparent retention of dry matter and crude protein compared with either the FFF- or FFF+ birds (P < 0.05). Birds fed FFF- diet had lower ceca digesta concentration of lactic acid than control and FFF+ (P < 0.05) birds. Results showed broiler breeder hens enriched egg yolk with n-3 polyunsaturated fatty acids without effects on the liver while the supplemental enzyme did not improve the utilization of FFF.

Multimodal Enzyme Delivery and Therapy Enabled by Cell Membrane-Coated Metal-Organic Framework Nanoparticles.

Therapeutic enzymes used for genetic disorders or metabolic diseases oftentimes suffer from suboptimal pharmacokinetics and stability. Nanodelivery systems have shown considerable promise for improving the performance of enzyme therapies. Here, we develop a cell membrane-camouflaged metal-organic framework (MOF) system with enhanced biocompatibility and functionality. The MOF core can efficiently encapsulate enzymes while maintaining their bioactivity. After the introduction of natural cell membrane coatings, the resulting nanoformulations can be safely administered in vivo. The surface receptors on the membrane can also provide additional functionalities that synergize with the encapsulated enzyme to target disease pathology from multiple dimensions. Employing uricase as a model enzyme, we demonstrate the utility of this approach in multiple animal disease models. The results support the use of cell membrane-coated MOFs for enzyme delivery, and this strategy could be leveraged to improve the usefulness of enzyme-based therapies for managing a wide range of important human health conditions.

Effect of supplementation of multiple enzymes to the diets containing variable protein sources on performance and nutrient utilization in commercial broilers.

The effect of supplementation of multiple enzymes at 0, 1X, and 2X concentration to the diet containing variable protein sources (replacement of soybean meal with 10% guar meal (GM), 10% rapeseed meal (RSM), and 10% cottonseed meal (CSM)) was studied in a factorial manner on performance, carcass yield, protein, and energy utilization in commercial broiler males. For this purpose, 600-day-old commercial male broiler chicks (Ross) were randomly divided into 12 treatment groups with 10 replications of 5 birds each and reared in battery brooders up to 42 days of age. Treatment groups are as follows: T1 control and T2 and T3 corn-soya diet with enzymes at 1X and 2X concentrations, respectively. From T4 to T12 treatments, corn-soya meal was replaced by 10% GM, RSM, and CSM without, with 1X and 2X concentration of enzyme supplementation, respectively. The cumulative body weight gain was significantly (P < 0.05) higher in control and birds fed with GM diet in pre-starter and starter as well as during overall period. Supplementation of multiple enzymes at 1X and 2X did not influence body weight, feed intake, and FCR (P > 0.05) during 0-42 days of age. During experimental period, birds fed with the control diet and 10% GM diet showed significantly (P < 0.05) higher cumulative feed intake and better FCR compared to birds fed with 10% RSM diet and 10% cottonseed meal (CSM). Supplementation of multiple enzymes did not influence the mortality and slaughter variables and protein utilization in broilers.

Dietitians' Evaluation of Clearing Luminal Occlusions of Gunk (DECLOG): A Pilot Feasibility Study.

BACKGROUND: Tube clogging is the most frequent mechanical complication associated with enteral nutrition. The objective of this study was to assess the efficacy of a protocol incorporating prophylactic use of a declogging system with enhanced patient education and monitoring to proactively reduce the incidence of tube occlusions in the home care setting. METHODS: A convenient sample of patients discharged from hospital to home enteral nutrition (HEN) was screened for eligibility and randomized to control group (standard care) or study group (standard care with prophylactic protocol and monitoring). Study patients received 4 enzyme declogging kits before discharge and were instructed to administer them every 7 days for 4 weeks. RESULTS: Seventeen of 49 (35%) patients reported tube occlusions. The incidence of tube occlusion in the control group was not statistically different than in the study group (29% vs 39%, P = 0.44). There was no difference between the 2 groups for negative tube outcomes, such as tube occlusion (P = 0.44), emergency department visit (P = 0.24), tube replacement (P = 0.99), or hospital readmission (P = 0.33). Continuous feeding method (P = 0.037), small-bowel feeding tubes (P = 0.016), and tube diameter <14 French (P = 0.069) were associated with tube occlusions. CONCLUSION: A prophylactic protocol using an enzyme declogging system did not lessen the likelihood of tube occlusions when compared with standard care. Multiple factors are associated with tube occlusion. More research investigating the use of a declogging system to prevent clogging incidence in patients receiving HEN is warranted.

The effectiveness of fibrolytic enzymes and active yeast on improving reticulorumen pH in dairy cows.

Exogenous fibrolytic enzymes (EFE) and yeast are feed supplements that improve forage digestion in rumen, but their influences on physical reticulorumen parameters are not well studied. This study was designed to evaluate the effect of the EFE:endo-ß-xylanase (37x104 U/cow/day), endocellulase (45x104 U/cow/day), endo-ß-glucanase (12x104U/cow/day), and active yeast - Saccharomyces cerevisiae CNCM-1077 (10x109CFU/cow/day) supplements on reticulorumen pH (RpH) and temperature (RT) in dairy cows. Nine Lithuanian Red cows were allocated into three groups (3 cows/group): control group (C) - farm diet without supplementation, enzyme group (E) - farm diet supplemented with EFE, enzyme and active yeast group (EY) - farm diet supplemented with EFE and active yeast. The feeding trial lasted for 60 d. All cows were equipped with reticuloruminal telemetric pH and temperature sensor device. Data provided by the device were used to calculate the mean RpH (RpH/24h), the mean minimal RpH (minRpH/24h) and mean of the time that RpH was below the threshold value of 6.0 (RpH⟨6.0/24h, min.). The highest RpH/24h (6.37±0.22) was observed in group EY and it was by 1.62% (p⟨0.05) and 1.27% (p⟨0.001) higher as compared with groups E and C, respectively. Also minRpH/24h (6.24±0.24) was highest in group EY and values were by 0.63% (p⟨0.001) and 0.65% (p⟨0.001) higher as compared with groups C and E, respectively. The shortest duration of RpH⟨6.0/24h, was recorded in group EY, and it was by 57.76% (p⟨0.05) and 47.87% shorter as compared with groups C and E, respectively. In conclusion, feed supplementation with EFE and Saccharomyces cerevisiae CNCM-1077 had beneficial effect on RpH.

Application of the Phage Lysin Ply5218 in the Treatment of Streptococcus suis Infection in Piglets.

Streptococcus suis (S. suis) is a gram-positive bacterium and zoonotic pathogen. Currently it poses a serious problem in the swine industry due to the emergence of antibiotic-resistant bacteria. Thus, novel antimicrobials against S. suis infections are urgently needed. In the previous study, a cell wall hydrolase or lysin derived from Streptococcus prophage phi5218, termed Ply5218, was identified. This lysin showed strong bacteriolytic activity against S. suis. In the current study, the in vitro data showed that after incubation with pig serum, the bacteriolytic efficacy of Ply5218 declined in a time-dependent manner. The in vivo assays indicated that a Ply5218 triple treatment (6, 24, and 48 h post infection) was effective against various serotypes of S. suis in a murine infection model. This regimen also alleviated streptococcal-induced clinical symptoms in piglets and significantly reduced the bacterial burden and levels of interleukin 6, a proinflammatory cytokine. This study indicates that Ply5218 shows strong antibacterial activity in pigs and has the potential to be used as a treatment for infectious diseases caused by S. suis.

Effects of in-feed enzymes on milk production and components, reproduction, and health in dairy cows.

Our objectives were to characterize responses in the field to a mix of fibrolytic enzymes using large commercial dairy herds and sufficient study power to evaluate milk production and reproductive responses to an enzyme treatment started during the precalving period. We hypothesized that the use of the enzyme treatment would increase milk production when provided to dairy cows precalving and for approximately 200 d of lactation. The study was conducted on 7,507 cows, in 8 replicates and 16 pens, at 3 dairies in the United States. Eight pens were randomly allocated as control pens and received no enzyme, and another 8 pens received enzyme treatment at a dose of 750 mL/t of dry matter feed. Milk production and energy-corrected milk yield were increased with the enzyme treatment by 0.70 and 0.80 kg/d, respectively, across a 5-month period. Milk fat percentage was not significantly increased by enzyme treatment, but milk fat yield was significantly increased by 0.040 kg/d, compared with controls. Milk protein yield increased 0.010 kg/d with enzyme treatment despite a small reduction of 0.020 percentage units in milk protein percentage. We found no evidence of an increase in the ln somatic cell count for the enzyme-treated cows. Body weight overall was not increased for enzyme-treated cows, but we did observe a numerical increase in dry matter intake (0.20 kg/head per day) for enzyme-treated cows. Most production responses to the enzyme treatment were influenced by dairy. Compared with controls, milk yield in enzyme-treated cows was significantly higher by 3.6 kg/d in dairy 2 and numerically higher by 0.60 and 0.20 kg/d in dairies 1 and 3, respectively. Reproduction, health, and risk of removal or death were not significantly influenced by treatment, apart from a reduced time to first breeding. Production responses to the enzyme treatment varied by dairy from substantial to minor increases, but variation among dairies was not evident in differences in dry matter intake or in partitioning of body weight among enzyme-treated and control pens and cows. It appears likely that the increase in production reflected increased digestibility of feed; however, further work is needed to identify factors influencing the variation in production responses to enzymes.

Digestibilidade do amido e comportamento ingestivo de novilhos confinados sob efeito de doses de complexo enzimático em dietas de alta densidade energética / Digestibility of starch and ingestive behavior of feedlot steers by effect of enzymatic complex doses in high-energy diets

O objetivo do presente trabalho foi avaliar a conversão alimentar, a digestibilidade do amido, o comportamento ingestivo e o escore de sobras da dieta e de fezes de novilhos confinados, suplementados com doses do complexo enzimático (0; 2,5; 5,0 e 7,5g animal-1 dia-1) e alimentados com dieta constituída por 85% de grão de milho e 15% de núcleo proteico, vitamínico e mineral, na base seca, isenta de forragem. O delineamento experimental foi o de blocos ao acaso contendo quatro tratamentos e quatro repetições. Trinta e dois novilhos inteiros, ½ sangue Angus Nelore, com idade média de 12 meses e peso vivo médio inicial de 422kg, foram confinados por um período de 77 dias. Cada grama de inclusão de complexo enzimático melhorou a conversão alimentar em 0,1652%, reduziu a matéria seca das fezes em 0,4648% e o tempo de ingestão de água em 0,0068 horas dia-1. A máxima digestibilidade do amido foi alcançada na dose de 5,08g animal-1 dia-1. A inclusão progressiva do complexo enzimático à dieta de alta densidade energética promoveu melhoria na conversão alimentar, redução na matéria seca das fezes e diminuição do tempo de ingestão de água. A dose de 5g animal-1 dia-1 do complexo enzimático aumentou a digestibilidade do amido.(AU)

The objective was to evaluate the feed conversion, starch digestibility, ingestive behavior, diet leftover score and feces score of steers supplemented with doses of enzyme complex (0; 2.5; 5.0 and 7 .5g animal -1 day -1 ) fed with roughage-free diet composed of a mixture of 85% whole corn grain and 15% protein-mineral-vitamin mix, on a dry matter basis. A completely randomized block design was adopted, consisting of four treatments and four replicates. Thirty-two ½ Angus ½ Nellore crossbred steers at an average age of 12 months, with an average initial weight of 422kg, were kept in a feedlot for 77 days. Each gram of enzyme complex inclusion improved feed conversion in 0.1652%, decreased feces dry matter in 0,4648% and time of water intake in 0.0068 hours day -1 . The maximum starch digestibility was reached in the dose of 5,08g animal -1 day -1 . The gradual inclusion of enzyme complex promoted improvement in feed conversion, reduction in the dry matter of feces and redution in the time of water intake. The enzyme complex dose of 5.0g animal -1 day -1 increased the starch digestibility.(AU)

Successes, failures, and future prospects of prodrugs and their clinical impact.

INTRODUCTION: Ample efforts have been carried out to improve the efficacy of a variety of drugs. The prodrugs approach was found to be a safe haven for providing medications with improved pharmacokinetic and pharmacodynamic properties. Areas covered: Herein, several selected successful prodrugs are reported and categorized. These include prodrugs for the treatment of the cardiovascular system, the central nervous system, the gastrointestinal tract, ophthalmology, the immune system, and oncology. In addition, some successful antiviral, antibacterial, antifungal, antiprotozoal, and several other miscellaneous prodrugs are documented. Further, a number of failed prodrugs are reported followed by those potentially promising prodrugs of the future. Expert opinion: The molecular revolution and accumulation of knowledge on the chemistry of enzymes and transporters has opened the door widely to novel successful prodrugs. For example, newer platelet aggregation inhibitors could signal the end of the warfarin era with their demanding treatment follow-up. The discovery of prodrugs can significantly improve the quality of patient care. Future attention should be focused towards directed enzyme prodrug therapy (DEPT). This strategy employs the design of artificial enzymes to activate prodrugs at specific sites. Agents designed for use in DEPT medicine can be directed at antibodies, genes, viruses, and clostridia.