Review: N1-methyl-pseudouridine (m1Ψ): Friend or foe of cancer?

Due to the health emergency created by SARS-CoV-2, the virus that causes the COVID-19 disease, the rapid implementation of a new vaccine technology was necessary. mRNA vaccines, being one of the cutting-edge new technologies, attracted significant interest and offered a lot of hope. The potential of these vaccines in preventing admission to hospitals and serious illness in people with comorbidities has recently been called into question due to the vaccines' rapidly waning immunity. Mounting evidence indicates that these vaccines, like many others, do not generate sterilizing immunity, leaving people vulnerable to recurrent infections. Additionally, it has been discovered that the mRNA vaccines inhibit essential immunological pathways, thus impairing early interferon signaling. Within the framework of COVID-19 vaccination, this inhibition ensures an appropriate spike protein synthesis and a reduced immune activation. Evidence is provided that adding 100 % of N1-methyl-pseudouridine (m1Ψ) to the mRNA vaccine in a melanoma model stimulated cancer growth and metastasis, while non-modified mRNA vaccines induced opposite results, thus suggesting that COVID-19 mRNA vaccines could aid cancer development. Based on this compelling evidence, we suggest that future clinical trials for cancers or infectious diseases should not use mRNA vaccines with a 100 % m1Ψ modification, but rather ones with the lower percentage of m1Ψ modification to avoid immune suppression.

Pelvic floor muscle length changes with breathing in males: A preliminary report.

The study aimed to identify whether pelvic floor muscles modulate length with breathing, and if any length changes induced by breathing relate to abdominal cavity displacement and intra-abdominal pressure. To investigate these relationships, displacement of pelvic landmarks that related to pelvic floor muscle length using transperineal ultrasound imaging, breath volume, intra-abdominal pressure, abdominal and ribcage displacement, and abdominal and anal sphincter muscle electromyography were measured during quiet breathing and breathing with increased dead-space in ten healthy men. Pelvic floor muscle landmark displacement modulated with ribcage motion during breathing. This relationship was stronger for: i) motion of the urethrovesical junction (puborectalis muscle length change) than the mid-urethra landmark (striated urethral sphincter muscle length change), and ii) dead-space breathing in standing than dead-space breathing in supine or quiet breathing in standing. In most (but not all) participants, the urethrovesical junction descended during inspiration and elevated during expiration. Striated urethral sphincter length changes during the respiratory cycle was independent of intra-abdominal pressure. In summary, breathing involves pelvic floor muscle length changes and is consistent with the role of these muscles during respiration to aid maintenance of continence, lung ventilation and/or provision of support to the abdominal cavity. Clinicians who train pelvic floor muscles need to be aware that length change of pelvic floor muscles is expected with breathing.

Evidence for increased tone or overactivity of pelvic floor muscles in pelvic health conditions: a systematic review.

OBJECTIVE: Pelvic floor muscle tone, which includes active and passive components, is argued to be increased in many pelvic health conditions, including those involving pain. This study systematically reviewed evidence for increased pelvic floor muscle tone in pelvic health conditions. DATA SOURCES: Electronic databases (PubMed, CINAHL, and Embase) were searched up to May 31, 2021. The search strategy included variants of pelvic and/or floor, muscle, and tone using keywords and Medical Subject Headings (MeSH) terms. STUDY ELIGIBILITY CRITERIA: Studies were included if they investigated increased tone of the pelvic floor muscle and reported measures of active or mechanical properties of the pelvic floor muscle in humans with any pelvic health condition, including pain, bowel, urogenital, or sexual dysfunctions. Studies of any design were included, except systematic and narrative reviews. Reference lists of studies, reviews, and book chapters were searched for additional studies. METHODS: Data were extracted using a standardized form, including measurement tool and outcome measure. Risk of bias was analyzed using a modified ROBINS-I (Risk of Bias In Non-randomized Studies - of Interventions) tool, and a score was allocated to determine whether the study provided "convincing" interpretation (comparison with condition-free control group, valid measure, no application issues). RESULTS: In total, 151 studies were included, reporting 8 different tools (electromyography, dynamometry, manometry, digital palpation, defecography, ultrasound, magnetic resonance imaging, other). The most common pelvic health condition was pelvic pain (n=16 conditions), followed by bowel and urogenital conditions. Most studies (57%) were cross-sectional. A healthy control group was infrequently included for comparison (27%). Unvalidated methods or methods applied in a manner that precluded convincing interpretation were common (94%). Of the 15 measurement tools that provided convincing evidence, 10 demonstrated greater tone in a pelvic health condition (all pain) compared with controls, and 5 showed no difference. CONCLUSION: Despite the large literature, few studies provide convincing evidence for increased tone/overactivity of pelvic floor muscles in pelvic health conditions. Interpretation is hampered by design and measurement issues. Terminology was often inaccurate. Few studies investigate male, transgender, and pediatric groups.

Nanosecond pulsed electric field treatment of human milk: Effects on microbiological inactivation, whey proteome and bioactive protein.

In the current study, nanosecond pulsed electric field (nsPEF) was investigated at lab-scale to optimise processing conditions of donor human milk to reduce bacterial counts, and to evaluate its effect on the bioactive proteins in human milk. Response surface methodology was utilized to optimise critical processing parameters. Two optimal nsPEF processing conditions were validated: 15 kV voltage, 6000 pulses at 20 Hz frequency, and 15 kV voltage, 6000 pulses at 50 Hz frequency. Compared to raw human milk, nsPEF processed milk had over 60 % retention of lysozyme, lactoperoxidase and lactoferrin, and 100 % retention of xanthine oxidase and immunoglobulin A. The contents of the five proteins were significantly higher after nsPEF processing when compared with Holder pasteurization. Liquid chromatography-mass spectrometry analysis showed that loss of milk proteins was smaller for samples treated with nsPEF than Holder pasteurization. These results indicated that nsPEF is a promising novel pasteurization method.

The Results of Different Heating Temperatures on Activities of Bioactive Proteins in Human Milk.

BACKGROUND: The most utilized pasteurization method in donor human milk banks is Holder pasteurization (heating 62.5 °C for 30 min). However, many bioactive proteins are heat sensitive and are inactivated. RESEARCH AIM: To determine the results of a range of heating regimes on the activities of xanthine oxidase, lactoperoxidase and lysozyme, the concentrations of immunoglobulin A and lactoferrin, as well as bacterial inactivation. METHOD: This prospective, cross-sectional, intervention study was designed to measure the influence of heating temperatures on bioactive components in donor human milk. Milk samples were processed at 40, 50, 55, 62.5, 75, 127 °C and the activities of the enzymes, and the concentration of immune proteins, were measured. RESULTS: No bacterial colonies were detectable, using standard culture methods, after heating above 50 ºC. All proteins studied retained over 60% concentrations or activities when the pasteurization temperature was 50 ºC or lower, while their concentrations or activities were lost at higher temperatures. For lactoferrin, the residual concentration was above 80% when heating temperature was under 55 °C, while only 20% remained after Holder pasteurization. Both xanthine oxidase and lactoperoxidase had little residual activity when temperatures were above Holder pasteurization. Lysozyme retained a greater proportion of residual activity than other proteins, following heating at all temperatures. CONCLUSIONS: The concentrations or activities of immune proteins and bioactive enzymes decreased when heated above 50 °C. The results of this study can be used to design temperature control guidance during alternative methods of pasteurization.

Comparative proteomic analysis of donor human milk pasteurized by hydrostatic high-pressure.

The aim of this study was to comprehensively investigate the effect of Holder pasteurization (HoP) compared with that of hydrostatic high-pressure (HHP) processing on human milk proteins, including milk fat globule membrane (MFGM) proteins, whey proteins and caseins. Milk fat globules in milk processed by high-pressure were similar to those in raw milk in terms of their size distribution and microstructure, while the globules in milk processed by HoP were aggregated. The protein profiles of milk subjected to HHP processing more closely resembled those of raw milk than HoP milk. Proteins in milk whey were less affected by HoP or HHP than MFGM and casein proteins. The findings indicated a better preservation of the protein profile for HHP compared to HoP of human milk.

Physicochemical Properties and Whey Proteomes of Camel Milk Powders Produced by Different Concentration and Dehydration Processes.

Camel milk powder production is an alternative to preserve the perishable milk for later-date consumption. However, the impacts of dehydration processes on bioactive compounds in camel milk are largely unknown. Hence, the present study attempted to compare the physicochemical properties and protein profiles of camel milk powders produced by different concentration and dehydration processes. Six camel milk powders were produced by freeze- and spray-drying methods in conjunction with two liquid concentration techniques, namely spray dewatering and reverse osmosis. The results of proteomic analysis showed that direct freeze-dried camel milk powder had the least changes in protein profile, followed by direct spray-dried powder. The camel milk powders that underwent concentration processes had more profound changes in their protein profiles. Among the bioactive proteins identified, lactotransferrin and oxidase/peroxidase had the most significant decreases in concentration following processing. On the contrary, glycosylation-dependent cell adhesion molecule 1, peptidoglycan recognition protein 1, and osteopontin increased in concentration. The results revealed that direct freeze drying was the most ideal method for preserving the bioactive proteins during camel milk powder production. However, the freeze-drying technique has cost and scalability constraints, and the current spray-drying technique needs improvement to better retain the bioactivity of camel milk during powder processing.

Comprehensive biochemical and proteomic characterization of seasonal Australian camel milk.

Although camel milk is increasingly becoming a popular alternative to bovine milk around the world including Australia, studies of Australian camel milk are still lacking. A comprehensive and systematic analysis of major nutritional components, physical properties, antimicrobial enzymes and whey proteomes of Australian camel milk obtained over four seasons was conducted, for the first time in present study. The composition and physical properties of Australian camel milk varied with season, milking frequency and yield. The highest lactoperoxidase and polyamine oxidase activity was observed in summer and winter, respectively. A total of 97 proteins were quantified, on a relative basis, across all the seasonal bulk milk samples. Summer camel milk contained higher amounts of functional whey proteins, such as lactotransferrin, peptidoglycan recognition protein 1, osteopontin and lactoperoxidase. These results contribute to a better understanding of the Australian camel milk and provide insights into processing of dairy products from this milk.

The influence of prostatectomy and body position on location and displacement of pelvic landmarks with pelvic floor muscle contraction.

AIMS: To compare pelvic floor muscle (PFM) anatomy and function (i) between pre- and post-prostatectomy in standing, and (ii) between sitting and standing postprostatectomy. METHODS: Thirty-two men scheduled to undergo a prostatectomy volunteered to participate. Transperineal ultrasound imaging was used to visualize five anatomical pelvic landmarks that have been validated to reflex anatomy and activity of PFMs (pubic symphysis, anorectal junction [ARJ], mid-urethra [MU], bulb of penis [BP], and urethrovesical junction [UVJ]). Both before and after prostatectomy, participants performed three submaximal PFM contractions in sitting and/or standing positions while ultrasound data were recorded. RESULTS: Postprostatectomy the UVJ location was more caudal and dorsal, the ARJ (puborectalis) vector was longer, the BP was more ventral than preprostatectomy, and these landmarks moved less ventrally with contraction. After prostatectomy, the MU, BP, and ARJ were more ventral in standing than sitting. The UVJ was more caudal and elevated more with contraction in standing than sitting after prostatectomy. CONCLUSION: These data demonstrate differences in the anatomy and mechanics of PFMs post- versus pre-prostatectomy, and between sitting and standing positions postprostatectomy. Findings are consistent with surgical changes to the bladder and urethral anatomy. Reduced passive support for the urethra and bladder are likely to may contribute to differences between standing and sitting postprostatectomy.

Comparing the effects of hydrostatic high-pressure processing vs holder pasteurisation on the microbial, biochemical and digestion properties of donor human milk.

In this study, hydrostatic high-pressure processing (HHP), a non-thermal pasteurisation method, was used to achieve the microbiological safety of donor human milk. After HHP, no bacteria were detected in human milk processed at 400 MPa for 5 min. Activities of a selection of bioactive components, including lysozyme, xanthine oxidase, lactoperoxidase, immunoglobulin A, lactoferrin, lipoprotein lipase and bile salt-stimulated lipase, did not decrease significantly. This study further investigated the gastrointestinal digestion kinetics of HoP and HHP milk compared with raw human milk, using an in vitro static infant digestion model. After 60 min of 'gastric digestion', the microstructure and protein profile of HHP milk samples were more similar to raw milk samples than HoP milk samples. Overall, HPP showed a better retention in milk nutrients and closer digestion behavior than that of HoP.

Digestibility of proteins in camel milk in comparison to bovine and human milk using an in vitro infant gastrointestinal digestion system.

The absence of ß-lactoglobulin, high ß-/αs-casein ratio and protective proteins make camel milk a promising alternative protein base for making human infant formulae. In this study, protein digestibility of camel milk was compared with that of bovine and human milk using an in vitro infant gastrointestinal digestion system. A low degree of gastric proteolysis was observed in all three kinds of milk, and a single clot was formed in camel milk. The soluble milk proteins remaining in the gastric digesta were digested rapidly and extensively in the intestinal phase, while the proteins in the camel milk clot were hydrolysed gradually. Despite several similarities, bioactive peptides unique to individual milk were identified in the three intestinal milk digesta. The results suggest that camel milk proteins are equally digestible as bovine and human milk proteins under infant gastrointestinal digestion conditions, and it may be a prospective substitute for infant formula base.

The repeatability of measurements of male pelvic floor anatomy and function made from transperineal ultrasound images of healthy men and those before and after prostatectomy.

AIMS: To investigate the inter- and intratester repeatability of measurement of the location and displacement of five pelvic landmarks related to pelvic floor muscles with transperineal ultrasound (TPUS) imaging recorded from healthy men and men before and after prostatectomy. METHODS: TPUS images were selected from four different participant groups: healthy men, men awaiting prostatectomy, men 2 weeks after prostatectomy, and men 12 months after prostatectomy. On two separate occasions, two assessors with different levels of experience performed analysis of location and displacement of five pelvic landmarks in images made at rest and during voluntary contraction. A two-way mixed effects, single measurement, absolute agreement intraclass correlation coefficient (ICC) was used to investigate the repeatability. RESULTS: Intertester reliability of all locations at rest for all groups was excellent (ICCs > 0.8) except for the craniocaudal coordinate of the ventral urethrovesical junction for men 2 weeks postprostatectomy and the anorectal junction for men with a cancerous prostate. Intertester reliability of the measurement of landmark displacement was acceptable (>0.5) for the dorsoventral axis of motion but not for the craniocaudal axis of motion for all landmarks across all groups. The more experienced assessor was consistently more repeatable. More deeply placed landmarks were more often excluded from analysis and had poorer reliability. CONCLUSIONS: Analysis of TPUS images across clinical groups is repeatable for both location and displacement of pelvic landmarks related to pelvic floor muscles when measures are made twice. Analysis experience, landmark depth and optimization of ultrasound settings appear to be important factors in reliability.

A sensitive and high-throughput fluorescent method for determination of oxidase activities in human, bovine, goat and camel milk.

Milk oxidases are an integral part of milk immune system, and good indicators for milk thermal history. Current assay methods for milk oxidases are either insensitive, tedious or not cost-effective. In this study, a high-throughput fluorescence assay method for determination of xanthine oxidase (XO) and polyamine oxidase (PAO) activities in milk samples was developed. The hydrogen peroxide generated by XO catalysed oxidation of hypoxanthine, and PAO catalysed oxidation of spermine, was coupled to horseradish peroxidase conversion of Amplex® Red (1-(3,7-dihydroxyphenoxazin-10-yl)ethanone) to the fluorescent product resorufin. The assay was highly sensitive, with limits of detection of activity in milk being 3 × 10-7 and 7 × 10-7 U/mL for XO and PAO, respectively. Intra-run and inter-run results showed good assay repeatability and reproducibility. The assay was successfully applied to survey the XO and PAO activities in human, bovine, goat and camel milk samples, and it can be readily adapted for measurements of other oxidase activities.

A sensitive, high-throughput fluorescent method for the determination of lactoperoxidase activities in milk and comparison in human, bovine, goat and camel milk.

Lactoperoxidase (LPO) is one of the major antibacterial ingredients in milk and an extensively employed indicator for milk heat treatment. The traditional method for LPO activity measurement using ABTS (2,2'-azinobis(3-ethylbenzothiazoline-6-sulphonate) cannot achieve high sensitivity and is affected by indigenous milk thiocyanate. A more sensitive microplate fluorescent assay was developed by monitoring generation of red-fluorescent resorufin from LPO catalysed oxidation of Amplex® Red (1-(3,7-dihydroxyphenoxazin-10-yl)ethanone) in this study. The assay is particularly suitable for milk LPO activity measurement as it eliminates the influences of indigenous milk hydrogen peroxide and thiocyanate. The method limit of detection was 7.1x10-6 U/mL of LPO in milk and good intra-run and inter-run precision was obtained. The LPO activities ranked as bovine > goat > camel > human in the four types of milk analysed. The high sensitivity and low cost of this assay makes it suitable for LPO activity analyses in both laboratory and commercial scales.

Sustainability management of short-lived freshwater fish in human-altered ecosystems should focus on adult survival.

Fish populations globally are susceptible to endangerment through exploitation and habitat loss. We present theoretical simulations to explore how reduced adult survival (age truncation) might affect short-lived freshwater fish species in human-altered contemporary environments. Our simulations evaluate two hypothetical "average fish" and five example fish species of age 1 or age 2 maturity. From a population equilibrium baseline representing a natural, unaltered environment we impose systematic reductions in adult survival and quantify how age truncation affects the causes of variation in population growth rate. We estimate the relative contributions to population growth rate arising from simulated temporal variation in age-specific vital rates and population structure. At equilibrium and irrespective of example species, population structure (first adult age class) and survival probability of the first two adult age classes are the most important determinants of population growth. As adult survival decreases, the first reproductive age class becomes increasingly important to variation in population growth. All simulated examples show the same general pattern of change with age truncation as known for exploited, longer-lived fish species in marine and freshwater environments. This implies age truncation is a general potential concern for fish biodiversity across life history strategies and ecosystems. Managers of short-lived, freshwater fishes in contemporary environments often focus on supporting reproduction to ensure population persistence. However, a strong focus on water management to support reproduction may reduce adult survival. Sustainability management needs a focus on mitigating adult mortality in human-altered ecosystems. A watershed spatial extent embracing land and water uses may be necessary to identify and mitigate causes of age truncation in freshwater species. Achieving higher adult survival will require paradigm transformations in society and government about water management priorities.

Influence of body position on dynamics of the pelvic floor measured with transperineal ultrasound imaging in men.

AIMS: This paper aims to evaluate the feasibility of transperineal ultrasound imaging (TPUS) for visualizing the motion of pelvic landmarks associated with striated pelvic floor muscle contraction in men in standing; to compare the locations of pelvic landmarks between sitting and standing; and to compare the effects of different body positions on measures of pelvic floor muscle contraction. METHODS: Thirty-five men awaiting prostatectomy volunteered to participate. Participants performed three repetitions of submaximal pelvic floor contraction in sitting and again in standing. Movement of pelvic landmarks with contraction was recorded using an ultrasound imaging transducer placed on the perineum. RESULTS: The feasibility of TPUS in men in standing was demonstrated through the visualization of three out of four pelvic landmarks in more than 95% of images in the standing position. Analysis of pelvic landmarks and their respective relationships with muscle shortening demonstrated that the anorectal junction and urethrovesical junction were lower and the estimated length of puborectalis was shorter in standing than sitting. The mid-urethra (striated urethral sphincter) and anorectal junction (puborectalis) landmark displaced further cranially in standing than sitting. CONCLUSIONS: TPUS can be used to visualize three pelvic landmarks in men with cancerous prostates. Puborectalis is shorter at rest in standing than sitting, and elevation of the mid-urethra and the anorectal junction is more in standing than sitting. Together these findings indicate that feedback for pelvic floor muscle training is possible in both positions, but the position needs to be standardized for a comparative assessment.

Preliminary Evidence for Enhanced Thymine Absorption: A Putative New Phenotype Associated With Fluoropyrimidine Toxicity in Cancer Patients.

BACKGROUND: Chemotherapy for colorectal, head and neck, and breast cancer continues to rely heavily on 5-fluorouracil and its oral prodrug capecitabine. Associations of serious fluoropyrimidine adverse effects have focused on inherited deficiency of the catabolic enzyme, dihydropyrimidine dehydrogenase. However, abnormal dihydropyrimidine dehydrogenase activity accounts for only about one-third of observed toxicity cases. Thus, the cause of most fluorouracil toxicity cases remains unexplained. METHODS: For this small cohort study, thymine (THY) 250 mg was administered orally to 6 patients who had experienced severe toxicity during treatment with 5FU or capecitabine. Plasma and urine were analyzed for THY and its catabolites dihydrothymine (DHT) and ß-ureidoisobutyrate. RESULTS: Of the 6 patients, 2 had decreased THY elimination and raised urinary THY recovery consistent with inherited partial dihydropyrimidine dehydrogenase deficiency, confirmed by DPYD sequencing. Unexpectedly, 3 patients displayed grossly raised plasma THY concentrations but normal elimination profiles (compared with a normal range for healthy volunteers previously published by the authors). DPYD and DPYS sequencing of these 3 patients did not reveal any significant loss-of-activity allelic variants. The authors labeled the phenotype in these 3 patients as "enhanced thymine absorption". Only 1 of the 6 cases of toxicity had a normal postdose plasma profile for THY and its catabolites. Postdose urine collections from all 6 patients had THY/DHT urinary ratios above 4.0, clearly separated from the ratios in healthy subjects that were all below 3.0. CONCLUSIONS: This small cohort provided evidence for a hypothesis that fluorouracil toxicity cases may include a previously undescribed pyrimidine absorption variant, "enhanced thymine absorption," and elevated THY/DHT ratios in urine may predict fluorouracil toxicity. A prospective study is currently being conducted.

De novo assembly of a tadpole shrimp (Triops newberryi) transcriptome and preliminary differential gene expression analysis.

Next-generation sequencing techniques, such as RNA sequencing, have provided a wealth of genomic information for nonmodel species. Transcriptomic information can be used to quantify the patterns of gene expression, which can identify how environmental differences invoke organismal stress responses and provide a gauge in predicting species adaptability. In our study, we used RNA sequencing to characterize the first transcriptome from a naupliar tadpole shrimp (Triops newberryi) to identify the genes expressed during the early life history stages and which could be important for future genomic studies. RNA was extracted from naupliar T. newberryi that were reared in a laboratory-controlled setting and in two different water types, a native and a non-native condition. A total of six replicates, three per condition, were sequenced with the Illumina Hi-Seq 2000 achieving 365 M 50-nt reads. High-quality reads were produced and de novo assembly was used to construct a T. newberryi transcriptome that was approximately 24.8 M base pairs. More than 10 000 peptides were predicted from the assembly, and genes were sorted into gene ontology categories. The use of different water conditions allowed for a preliminary differential gene expression analysis in order to compare the changes in gene expression between conditions. There were 299 differentially expressed genes between water conditions that might serve as a focal point for future genomic studies of Triops acclimation to different environments. The Triops transcriptome could serve as vital genomic information for additional studies on Branchiopod crustaceans.

Plasma sodium measurements by direct ion selective methods in laboratory and point of care may not be clinically interchangeable.

An estimated 25 % of indirect ion selective electrode (ISE) ICU plasma sodium measurements differ from corresponding direct ISE values by at least 4 mmol/L, the dominant factor being indirect ISE over-estimation driven by hypoproteinemia. Since direct measurements are considered unaffected by protein concentrations, we investigated whether direct ISE plasma sodium measurements in the laboratory and at point of care in ICU show sufficient agreement to be clinically interchangeable. From a 5 year clinical chemistry database, 9910 ICU plasma samples were assessed for agreement between direct ISE sodium measurements in ICU (ABL 700) and in the central laboratory (Vitros Fusion). The relationship between differences in paired plasma sodium measurements (Vitros-ABL) and total plasma protein concentrations was evaluated by generalized estimating equation linear regression. Patients were hypo-proteinemic [mean (SD) total protein concentration 56.9 (9.04) g/L]. Mean (SD) paired Vitros-ABL sodium measurements was -0.087 (1.74) mmol/L, range -14 to +10 mmol/L. Disagreement at ≥|4|mmol/L, ≥|3|mmol/L and ≥|2|mmol/L was present in 409 (4.1 %), 1333 (13.4 %) and 3591 (36.2 %) pairs respectively. Test-retest disagreement estimates within either source alone were substantially lower. Small negative Vitros-ABL differences associated with low plasma protein concentrations were reversed at high protein concentrations. Disagreement between plasma sodium concentrations monitored by two common direct ISE analyzers was substantially less than reported between direct and indirect ISE devices, although a protein influence of low clinical importance persisted. Disagreement was sufficient to jeopardize safe interchangeable interpretation in situations with a low tolerance for imprecision, such as hyponatremia correction.