Piglet gut microbial shifts early in life:causes and effects

The gut microbiome has long been known to play fundamentally important roles in the animal health and the well-being of its host. As such, the establishment and maintenance of a beneficial gut microbiota early in life is crucial in pigs, since early gut colonizers are pivotal in the establishment of permanent microbial community structures affecting the health and growth performance of pigs later in life. Emphasizing this importance of early gut colonizers, it is critical to understand the factors impacting the establishment of the piglet gut microbiome at weaning. Factors include, among others, diet, in-feed antibiotics, probiotics and prebiotic administration. The impact of these factors on establishment of the gut microbiome of piglets at weaning includes effects on piglet gut microbial diversity, structure, and succession. In this review, we thoroughly reviewed the most recent findings on the piglet gut microbiome shifts as influenced by weaning, and how these microbiome changes brought about by various factors that have been shown to affect the development of microbiota in piglets. This review will provide a general overview of recent studies that can help to facilitate the design of new strategies to modulate the gut microbiome in order to enhance gastrointestinal health, growth performance and well-being of piglets.

Less common etiologies of exocrine pancreatic insufficiency

Exocrine pancreatic insufficiency(EPI), an important cause of maldigestion and malabsorption, results from primary pancreatic diseases or secondarily impaired exocrine pancreatic function. Besides cystic fibrosis and chronic pancreatitis, the most common etiologies of EPI, other causes of EPI include unresectable pancreatic cancer, metabolic diseases(diabetes); impaired hormonal stimulation of exocrine pancreatic secretion by cholecystokinin(CCK); celiac or inflammatory bowel disease(IBD) due to loss of intestinal brush border proteins; and gastrointestinal surgery(asynchrony between motor and secretory functions, impaired enteropancreatic feedback, and inadequate mixing of pancreatic secretions with food). This paper reviews such conditions that have less straightforward associations with EPI and examines the role of pancreatic enzyme replacement therapy(PERT). Relevant literature was identified by database searches. Most patients with inoperable pancreatic cancer develop EPI(66%-92%). EPI occurs in patients with type 1(26%-57%) or type 2 diabetes(20%-36%) and is typically mild to moderate; by definition, all patients with type 3 c(pancreatogenic) diabetes have EPI. EPI occurs in untreated celiac disease(4%-80%), but typically resolves on a gluten-free diet. EPI manifests in patients with IBD(14%-74%) and up to 100% of gastrointestinal surgery patients(47%-100%; dependent on surgical site). With the paucity of published studies on PERT use for these conditions, recommendations for or against PERT use remain ambiguous. The authors conclude that there is an urgent need to conduct robust clinical studies to understand the validity and nature of associations between EPI and medical conditions beyond those with proven mechanisms, and examine the potential role for PERT.

The dynamics of the piglet gut microbiome during the weaning transition in association with health and nutrition

Background: Understanding the composition of the microbial community and its functional capacity during weaning is important for pig production as bacteria play important roles in the pig’s health and growth performance. However,limited information is available regarding the composition and function of the gut microbiome of piglets in early-life.Therefore, we performed 16 S rRNA gene and whole metagenome shotgun sequencing of DNA from fecal samples from healthy piglets during weaning to measure microbiome shifts, and to identify the potential contribution of the early-life microbiota in shaping piglet health with a focus on microbial stress responses, carbohydrate and amino acid metabolism.Results: The analysis of 16 S rR NA genes and whole metagenome shotgun sequencing revealed significant compositional and functional differences between the fecal microbiome in nursing and weaned piglets. The fecal microbiome of the nursing piglets showed higher relative abundance of bacteria in the genus Bacteroides with abundant gene families related to the utilization of lactose and galactose. Prevotel a and Lactobacil us were enriched in weaned piglets with an enrichment for the gene families associated with carbohydrate and amino acid metabolism. In addition, an analysis of the functional capacity of the fecal microbiome showed higher abundances of genes associated with heat shock and oxidative stress in the metagenome of weaned piglets compared to nursing piglets.Conclusions: Overal, our data show that microbial shifts and changes in functional capacities of the piglet fecal microbiome resulted in potential reductions in the effects of stress, including dietary changes that occur during weaning.These results provide us with new insights into the piglet gut microbiome that contributes to the growth of the animal.

Development of a rapid GC-FID method to simultaneously determine triethylamine, diisopropylamine, and 1,1,3,3-tetramethylguanidine residues in an active pharmaceutical ingredient

A rapid GC-FID method was developed to simultaneously determine residual levels of triethylamine(TEA), 1,1,3,3-tetramethylguanidine(TMG), and diisopropylamine(DIPA) in the synthetic route of an active pharmaceutical ingredient(API). Due to the severe absorption of amines on GC stationary phases,GC columns with various stationary phases were evaluated for optimal peak shape and reproducibility.The final conditions used the Agilent CP-Volamine column to resolve the three amines in 12 min. Various inlet liners were also screened to further improve the sensitivity of the analysis. The Restek Siltek~? liner was selected to achieve the desired detectability for the method. The quantitation limits were 4, 3, and 4 mg/mL for TEA, DIPA, and TMG in the presence of API, respectively. All three amines showed good linearity(r > 0.999) and recoveries(> 90%) over the concentration range of 3 to 16 mg/mL. The testing of residual amines was initially performed at the penultimate stage of the synthesis. However, this work demonstrates that TMG can act as a proton sponge to react with salicylic acid, the counter ion of the penultimate, to form a volatile component that elutes at a different retention time. Consequently, in the final method, these three amines were monitored in the final API to circumvent the matrix interference.Key parameters of the method were qualified per method validation requirements in ICH guidelines. The method was successfully applied for batch testing during development and implemented as an inprocess control procedure at manufacturing sites.

Strategies for PET imaging of the receptor for advanced glycation endproducts(RAGE)

The implication of the receptor for advanced glycation end-products(RAGE)in numerous diseases and neurodegenerative disorders makes it interesting both as a therapeutic target and as an inflammatory biomarker.In the context of investigating RAGE as a biomarker,there is interest in developing radiotracers that will enable quantification of RAGE using positron emission tomography(PET)imaging.We have synthesized potential small molecule radiotracers for both the intracellular([18F]InRAGER)and extracellular([18F]RAGER)domains of RAGE.Herein we report preclinical evaluation of both using in vitro(lead panel screens)and in vivo(rodent and nonhuman primate PET imaging)methods.Both radiotracers have high affinity for RAGE and show good brain uptake,but suffer from off-target binding.The source of the off-target PET signal is not attributable to binding to melatonin receptors,but remains unexplained.We have also investigated use of lipopolysaccharide(LPS)-treated mice as a possible animal model with upregulated RAGE for evaluation of new imaging agents.Immunoreactivity of the mouse brain sections revealed increases in RAGE in the male cohorts,but no difference in the female groups.However,it proves challenging to quantify the changes in RAGE due to off-target binding of the radiotracers.Nevertheless,they are appropriate lead scaffolds for future development of 2nd generation RAGE PET radiotracers because of their high affinity for the receptor and good CNS penetration.

Impact of adalimumab on disease burden in moderate-to-severe ulcerative colitis patients: The one-year, real-world UCanADA study

BACKGROUND A gap remains in documenting the impact of anti-tumor necrosis factor therapy on disease burden in ulcerative colitis(UC)patients treated in a real-world setting.The use of patient-reported outcomes(PROs)has been discussed as a primary endpoint in the context of the FDA PRO Guidance,for labelling purposes.Specifically,the efficacy and safety of adalimumab have been demonstrated in pivotal trials;however,data are needed to understand how clinical results translate into improvements in key aspects of the daily lives of UC patients,such as symptoms,health-related quality of life(HRQoL),and disability.AIM To assess real-world effectiveness of adalimumab on PRO measures in patients with moderate-to-severe UC.METHODS UCanADA was a single arm,prospective,1-year multicenter Canadian post-marketing observational study in which multiple PRO questionnaires were completed—with psychologic distress/depression symptoms as the primary endpoint—by patients with moderate-to-severe UC.Assessments were performed during patients’routine care visit schedule,which was at the initiation of adalimumab(baseline),after induction(approximately 8 wk),and 52 wk after baseline.Additional optional assessments between weeks 8 and 52 were collected at least once but no more than two times during this period.Serious safety events and per-protocol adverse events were collected.RESULTS From 23 Canadian centres,100 patients were enrolled and 48 completed the study.Measured with the Patient Health Questionnaire–9 items at week 52,61.5%(40/65)[95%confidence interval(CI):49.7%-73.4%]of the patients improved in psychologic distress/depression symptoms,which was slightly higher in completers[65.9%(29/44);95%CI:51.9%-79.9%].At week 52,clinical response and clinical remission were achieved respectively by 65.7%(44/73)and 47.8%(32/73)of the patients.The odds of improving depressive symptoms for those achieving a clinical remission at week 52 was 7.94 higher compared with those not achieving a clinical remission(CI:1.42,44.41;P=0.018).Significant changes from baseline to weeks 8 and 52 were observed in disability,HRQoL,and fatigue.Meaningful improvement was reported in work impairment.CONCLUSION At week 52,over 60%of the UCanADA patients had depressive symptoms significantly reduced,as well as HRQoL,fatigue symptoms,and work impairment improved.No new safety signals were detected.

Tumor-intrinsic metabolic reprogramming and how it drives resistance to anti-PD-1/PD-L1 treatment

The development of immune checkpoint blockade (ICB) therapies has been instrumental in advancing the field of immunotherapy. Despite the prominence of these treatments, many patients exhibit primary or acquired resistance, rendering them ineffective. For example, anti-programmed cell death protein 1 (anti-PD-1)/anti-programmed cell death ligand 1 (anti-PD-L1) treatments are widely utilized across a range of cancer indications, but the response rate is only 10%-30%. As such, it is necessary for researchers to identify targets and develop drugs that can be used in combination with existing ICB therapies to overcome resistance. The intersection of cancer, metabolism, and the immune system has gained considerable traction in recent years as a way to comprehensively study the mechanisms that drive oncogenesis, immune evasion, and immunotherapy resistance. As a result, new research is continuously emerging in support of targeting metabolic pathways as an adjuvant to ICB to boost patient response and overcome resistance. Due to the plethora of studies in recent years highlighting this notion, this review will integrate the relevant articles that demonstrate how tumor-derived alterations in energy, amino acid, and lipid metabolism dysregulate anti-tumor immune responses and drive resistance to anti-PD-1/PD-L1 therapy.

在初治和经治丙型肝炎病毒基因1b型慢性感染的代偿期肝硬化亚洲成年患者中评价奥比帕利和达塞布韦联合利巴韦林治疗的有效性和安全性

目的 评估奥比帕利（奥比他韦/帕立瑞韦/利托那韦）25/150/100mg,1次/d和达塞布韦（DSV） 250 mg,2次/d联合利巴韦林在丙型肝炎病毒基因1b型感染的代偿期肝硬化中国大陆成年患者中的有效性和安全性. 方法 采用开放标签、多中心Ⅲ期临床试验,在中国大陆、中国台湾地区和韩国开展,纳入初治和经治基因1b型丙型肝炎病毒感染的代偿期肝硬化（Metavir 评分纤维化分期=F4）成年患者,接受奥比他韦/帕立瑞韦/利托那韦和达塞布韦联合利巴韦林治疗12周.评估患者停药12周获得的持续病毒学应答（SVR）及24周获得的SVR率,并在接受至少1次研究药物的患者中评估有效性和安全性. 结果 共纳入63例中国大陆患者,其中62例（98.4％）患者基线Child-Pugh评分为5分.患者总体SVR12率及SVR24率为100％ （95％ CI：94.3％ ～ 100.0％）.发生的大多数不良事件为轻度.常见（发生率≥10％）所有级别不良事件和实验室异常包括总胆红素升高（36.5％）、乏力（19.0％）、非结合胆红素升高（19.0％）、结合胆红素升高（17.5％）和贫血（14.3％）.3例（4.8％）患者发生≥3级不良事件,均被研究者判定为与研究药物无关.无患者出现导致提前停药的不良事件. 结论 丙型肝炎病毒基因1b型感染的代偿期肝硬化中国大陆患者接受奥比他韦/帕立瑞韦/利托那韦和达塞布韦联合利巴韦林治疗12周,停药12周和24周的SVR均为100％.耐受性及安全性良好,大多数不良事件为轻度.

在初治和经治丙型肝炎病毒基因1b型慢性感染的非肝硬化亚洲成年患者中评价奥比帕利联合达塞布韦治疗的有效性和安全性：随机、双盲、安慰剂对照研究

目的 评估奥比帕利（奥比他韦/帕立瑞韦/利托那韦,OBV/PTV/r）25/150/100mg,1次/d,联合达塞布韦250mg,2次/d,在丙型肝炎病毒基因1b型感染的初治和经治非肝硬化中国成年患者中的有效性和安全性.方法 采用随机、双盲、安慰剂对照、多中心Ⅲ期临床试验,在中国大陆、韩国、中国台湾地区进行.纳入基因1b型初治和普通干扰素/聚乙二醇干扰素α联合利巴韦林经治非肝硬化患者,随机分为2组,分别接受OBV/PTV/r联合达塞布韦方案立即治疗12周（A组）,或接受安慰剂治疗12周后再继续接受OBV/PTV/r联合达塞布韦方案治疗12周（B组）.评估患者停药12周获得的持续病毒学应答（SVR12）及停药24周获得的持续病毒学应答（SVR24）率,及双盲阶段和开放标签阶段治疗后的不良事件和实验室异常发生率. 结果 共纳入410例中国大陆患者资料,按1∶1比例随机分至A组和B组（每组205例）.A组（205例）中初治患者的SVR12及SVR24率均为99％ （95％CI：94.8％ ～ 99.8％）,经治患者的SVR12及SVR24率均为100％ （95％ CI：96.3％～ 100％）,不同基线特征对SVR12及SVR24率没有影响.发生的大多数不良事件为轻度,治疗期间≥3级实验室异常少见,主要包括丙氨酸氨基转移酶升高（双盲阶段A组2例）、天冬氨酸氨基转移酶升高（双盲阶段A组3例）和总胆红素升高（开放标签阶段B组1例）,通常无症状,用药中断或停药后可恢复.仅有1例因不良事件停药（B组,开放标签阶段）. 结论 疗程为12周的OBV/PTV/r联合达塞布韦方案治疗中国基因1b型初治和经治非肝硬化慢性丙型肝炎患者可获得99％～ 100％的SVR12及SVR24率,且耐受性和安全性良好.

Venetoclax联合低剂量阿糖胞苷治疗不耐受强化化疗的初治急性髓系白血病患者:一项Ⅲ期随机、安慰剂对照试验中中国队列结果

目的:探讨Venetoclax联合低剂量阿糖胞苷(LDAC)在不能耐受强化诱导化疗的中国急性髓系白血病(AML)患者中的疗效及安全性。方法:一项Ⅲ期随机、双盲安慰剂对照试验(VIALE-C)中中国队列的结果。在本项国际临床试验中,入组了不适合接受强化化疗、新诊断为AML的18岁或以上的成人患者。在全球范围内,患者(211例)按2∶1的比例随机分配接受Venetoclax+LDAC或安慰剂+LDAC(28 d为1个周期),在第1~10天接受LDAC。主要研究终点为总生存(OS);次要研究终点包括缓解率、无事件生存期及不良事件(AE)。结果:入组15例中国患者(Venetoclax组9例;安慰剂组6例)。中位年龄为72(61~86)岁。与安慰剂组相比,Venetoclax组的死亡风险下降38%(HR=0.62,95%CI 0.12~3.07)。对延长6个月随访进行的计划外分析显示,Venetoclax组的中位OS时间为9.0个月,安慰剂组为4.1个月。完全缓解(CR)率与血细胞计数未完全恢复的CR(CRi)率分别为33%(3/9)和0(0/6)。最常见的非血液学AE(Venetoclax组与安慰剂组)为低钾血症(5/9和4/6)、呕吐(4/9和3/6)、便秘(2/9和4/6)和低白蛋白血症(1/9和4/6)。结论:Venetoclax联合LDAC在中国患者中表现出有意义的疗效和可管理的安全性特征,这与在全球VIALE-C人群中的观察结果一致,使其成为不适合接受强化化疗的新诊断AML患者的一个重要治疗选择。

Potent and conditional redirected T cell killing of tumor cells using Half DVD-Ig

Novel biologics that redirect cytotoxic T lymphocytes （CTLs） to kill tumor cells bearing a tumor associated antigen hold great promise in the clinic. However, the ability to safely and potently target CD3 on CTL toward tumor associated antigens （TAA） expressed on tumor cells remains a challenge of both technology and biol- ogy. Herein we describe the use of a Half DVD-Ig format that can redirect CTL to kill tumor cells. Notably, Half DVD-Ig molecules that are monovalent for each speci- ficity demonstrated reduced non-specific CTL activation and conditional CTL activation upon binding to TAA compared to intact tetravalent DVD-Ig molecules that are bivalent for each specificity, while maintaining good drug like properties and appropriate PK properties.

Activation of RAS/MAPK pathway confers MCL-1 mediated acquired resistance to BCL-2 inhibitor venetoclax in acute myeloid leukemia

Despite high initial response rates,acute myeloid leukemia(AML)treated with the BCL-2-selective inhibitor venetoclax(VEN)alone or in combinations commonly acquires resistance.We performed gene/protein expression,metabolomic and methylation analyses of isogenic AML cell lines sensitive or resistant to VEN,and identified the activation of RAS/MAPK pathway,leading to increased stability and higher levels of MCL-1 protein,as a major acquired mechanism of VEN resistance.

Role of cellular reprogramming and epigenetic dysregulation in acquired chemoresistance in breast cancer

Acquired resistance to chemotherapy is a major limitation in clinical treatment for breast cancer.Accumulating evidence from in vitro,in vivo and clinical studies suggest that acquired chemoresistance is progressive,multifactorial and involve genetic and epigenetic aberrations.Among various mechanisms that contribute to chemoresistance,cellular reprogramming has extensively been implicated in breast cancer resistance lately.Cellular reprogramming events such as acquisition of epithelial to mesenchymal transition(EMT)and cancer stemness(CSCs)not only provide cancer cells with reversible phenotypic plasticity and survival advantage against cytotoxicity but also leads to aggressiveness,metastasis,clinical resistance,tumor recurrence and poor survival.The transient and reversible nature of cellular reprogramming processes and their controlled interaction with epigenetic regulatory complexes strongly support the involvement of dynamic epigenetic regulatory network in governing the cellular reprogramming and associated acquired chemoresistance.Further,epigenetic modulations are also gaining interest as promising interventions addressing the cancer cell reprogramming machinery to overcome acquired chemoresistance.This review discusses the previous reports and our recent findings that lead to current understanding of epigenetic dysregulation dictating the cellular reprogramming processes such as acquisition of EMT and CSCs phenotype and how they co-ordinate to establish acquired drug resistance in breast cancer.

Redefinition of Fatty Liver Disease from NAFLD to MAFLD through the Lens of Drug Development and Regulatory Science

Metabolic(dysfunction)-associated fatty liver disease(MAFLD)affects a third of the population and is a leading cause of liver-related death.Since no effective treatments exist,novel approaches to drug development are required.Unfortunately,outdated terminology and definitions of the disease are hampering efforts to develop new drugs and treatments.An international consensus panel has put forth an influential proposal for the disease to be renamed from nonalcoholic fatty liver disease(NAFLD)to MAFLD,includ-ing a proposal for how the disease should be diagnosed.As allies with the many stakeholders in MAFLD care―including patients,patients’advocates,clinicians,researchers,nurse and allied health groups,regional societies,and others―we are aware of the negative consequences of the NAFLD term and definition.We share the sense of urgency for change and will act in new ways to achieve our goals.Although there is much work to be done to overcome clinical inertia and reverse worrisome recent trends,the MAFLD initiative provides a firm foundation to build on.It provides a roadmap for moving for-ward toward more efficient care and affordable,sustainable drug and device innovation in MAFLD care.We hope it will bring promising new opportunities for a brighter future for MAFLD care and improve care and outcomes for patients of one of the globe’s largest and costliest public health burdens.From this viewpoint,we have revisited this initiative through the perspectives of drug development and regulatory science.