Modeling multiphase fluid flow,mass transfer,and chemical reactions in bioreactors using large-eddy simulation

We present a transient large eddy simulation (LES) modeling approach for simulating the interlinked physics describing free surface hydrodynamics, multiphase mixing, reaction kinetics, and mass transport in bioreactor systems. Presented case-studies include non-reacting and reacting bioreactor systems, modeled through the inclusion of uniform reaction rates and more complex biochemical reactions described using Contois type kinetics. It is shown that the presence of reactions can result in a non-uniform spatially varying species concentration field, the magnitude and extent of which is directly related to the reaction rates and the underlying variations in the local volumetric mass transfer coefficient.     

Evaluating the Arrhenius equation for developmental processes

The famous Arrhenius equation is well suited to describing the temperature dependence of chemical reactions but has also been used for complicated biological processes. Here, we evaluate how well the simple Arrhenius equation predicts complex multi‐step biological processes, using frog and fruit fly embryogenesis as two canonical models. We find that the Arrhenius equation provides a good approximation for the temperature dependence of embryogenesis, even though individual developmental intervals scale differently with temperature. At low and high temperatures, however, we observed significant departures from idealized Arrhenius Law behavior. When we model multi‐step reactions of idealized chemical networks, we are unable to generate comparable deviations from linearity. In contrast, we find the two enzymes GAPDH and β‐galactosidase show non‐linearity in the Arrhenius plot similar to our observations of embryonic development. Thus, we find that complex embryonic development can be well approximated by the simple Arrhenius equation regardless of non‐uniform developmental scaling and propose that the observed departure from this law likely results more from non‐idealized individual steps rather than from the complexity of the system.     

Biological and molecular profile of fracture non‐union tissue: A systematic review and an update on current insights

Fracture non‐union represents a common complication, seen in 5%–10% of all acute fractures. Despite the enhancement in scientific understanding and treatment methods, rates of fracture non‐union remain largely unchanged over the years. This systematic review investigates the biological, molecular and genetic profiles of both (i) non‐union tissue and (ii) non–union‐related tissues, and the genetic predisposition to fracture non‐union. This is crucially important as it could facilitate earlier identification and targeted treatment of high‐risk patients, along with improving our understanding on pathophysiology of fracture non‐union. Since this is an update on our previous systematic review, we searched the literature indexed in PubMed Medline; Ovid Medline; Embase; Scopus; Google Scholar; and the Cochrane Library using Medical Subject Heading (MeSH) or Title/Abstract words (non‐union(s), non‐union(s), human, tissue, bone morphogenic protein(s) (BMPs) and MSCs) from August 2014 (date of our previous publication) to 2 October 2021 for non‐union tissue studies, whereas no date restrictions imposed on non–union‐related tissue studies. Inclusion criteria of this systematic review are human studies investigating the characteristics and properties of non‐union tissue and non–union‐related tissues, available in full‐text English language. Limitations of this systematic review are exclusion of animal studies, the heterogeneity in the definition of non‐union and timing of tissue harvest seen in the included studies, and the search term MSC which may result in the exclusion of studies using historical terms such as ‘osteoprogenitors’ and ‘skeletal stem cells’. A total of 24 studies (non‐union tissue: n = 10; non–union‐related tissues: n = 14) met the inclusion criteria. Soft tissue interposition, bony sclerosis of fracture ends and complete obliteration of medullary canal are commonest macroscopic appearances of non‐unions. Non‐union tissue colour and surrounding fluid are two important characteristics that could be used clinically to distinguish between septic and aseptic non‐unions. Atrophic non‐unions had a predominance of endochondral bone formation and lower cellular density, when compared against hypertrophic non‐unions. Vascular tissues were present in both atrophic and hypertrophic non‐unions, with no difference in vessel density between the two. Studies have found non‐union tissue to contain biologically active MSCs with potential for osteoblastic, chondrogenic and adipogenic differentiation. Proliferative capacity of non‐union tissue MSCs was comparable to that of bone marrow MSCs. Rates of cell senescence of non‐union tissue remain inconclusive and require further investigation. There was a lower BMP expression in non‐union site and absent in the extracellular matrix, with no difference observed between atrophic and hypertrophic non‐unions. The reduced BMP‐7 gene expression and elevated levels of its inhibitors (Chordin, Noggin and Gremlin) could potentially explain impaired bone healing observed in non‐union MSCs. Expression of Dkk‐1 in osteogenic medium was higher in non‐union MSCs. Numerous genetic polymorphisms associated with fracture non‐union have been identified, with some involving the BMP and MMP pathways. Further research is required on determining the sensitivity and specificity of molecular and genetic profiling of relevant tissues as a potential screening biomarker for fracture non‐unions.     

From coarse to fine: the absolute Escherichia coli proteome under diverse growth conditions

Accurate measurements of cellular protein concentrations are invaluable to quantitative studies of gene expression and physiology in living cells. Here, we developed a versatile mass spectrometric workflow based on data‐independent acquisition proteomics (DIA/SWATH) together with a novel protein inference algorithm (xTop). We used this workflow to accurately quantify absolute protein abundances in Escherichia coli for > 2,000 proteins over > 60 growth conditions, including nutrient limitations, non‐metabolic stresses, and non‐planktonic states. The resulting high‐quality dataset of protein mass fractions allowed us to characterize proteome responses from a coarse (groups of related proteins) to a fine (individual) protein level. Hereby, a plethora of novel biological findings could be elucidated, including the generic upregulation of low‐abundant proteins under various metabolic limitations, the non‐specificity of catabolic enzymes upregulated under carbon limitation, the lack of large‐scale proteome reallocation under stress compared to nutrient limitations, as well as surprising strain‐dependent effects important for biofilm formation. These results present valuable resources for the systems biology community and can be used for future multi‐omics studies of gene regulation and metabolic control in E. coli.     

Match and mismatch: Integrating consumptive effects of predators,prey traits,and habitat selection in colonizing aquatic insects

Predators are a particularly critical component of habitat quality, as they affect survival, morphology, behavior, population size, and community structure through both consumptive and non‐consumptive effects. Non‐consumptive effects can often exceed consumptive effects, but their relative importance is undetermined in many systems. Our objective was to determine the consumptive and non‐consumptive effects of a predaceous aquatic insect, Notonecta irrorata, on colonizing aquatic beetles. We tested how N. irrorata affected survival and habitat selection of colonizing aquatic beetles, how beetle traits contributed to their vulnerability to predation by N. irrorata, and how combined consumptive and non‐consumptive effects affected populations and community structure. Predation vulnerabilities ranged from 0% to 95% mortality, with size, swimming, and exoskeleton traits generating species‐specific vulnerabilities. Habitat selection ranged from predator avoidance to preferentially colonizing predator patches. Attraction of Dytiscidae to N. irrorata may be a natural ecological trap given similar cues produced by these taxa. Hence, species‐specific habitat selection by prey can be either predator‐avoidance responses that reduce consumptive effects, or responses that magnify predator effects. Notonecta irrorata had both strong consumptive and non‐consumptive effects on populations and communities, while combined effects predicted even more distinct communities and populations across patches with or without predators. Our results illustrate that an aquatic invertebrate predator can have functionally unique consumptive effects on prey, attracting and repelling prey, while prey have functionally unique responses to predators. Determining species‐specific consumptive and non‐consumptive effects is important to understand patterns of species diversity across landscapes.     

Salivary non‐apoptotic tumoral microvesicles: A potential progressive marker in oral cancer patients

Tumour cell‐secreted microvesicles (MVs) contribute immensely to tumour progression. However, the role of tumoral salivary MVs in oral squamous cell carcinoma (OSCC) remains unclear. Herein, we elucidated the role of non‐apoptotic salivary tumoral MVs in OSCC development, especially relating to the migration ability. We purified and compared non‐apoptotic salivary tumoral MVs from 63 OSCC patients and orthotopic OSCC mice model. Next, we compared the protein difference between apoptotic and non‐apoptotic MVs by Western blot, proteomics and flow cytometry from saliva and CAL27 cells. Finally, we collected the non‐apoptotic MVs and co‐cultured with normal oral epithelial cells, the migration ability was examined by wound healing assay and Western blot assay. Our results indicated that the levels of non‐apoptotic tumoral S‐MVs were significantly higher in OSCC patients with T3 to T4 stages than in patients with T1 to T2 stages or healthy donors. In OSCC mice model, we found elevations of non‐apoptotic tumoral MVs associated with tumoral volume. EGFR overexpression increased the generation of non‐apoptotic tumoral MVs which could significantly promote normal epithelial cell migration. In conclusion, elevated levels of non‐apoptotic tumoral S‐MVs are associated with clinicopathologic features of OSCC patients, implying that non‐apoptotic tumoral S‐MVs are a potential progressive marker of OSCC.     

Comparison of native and non‐native predator consumption rates and prey avoidance behavior in North America and Europe

Novel predator–prey interactions can contribute to the invasion success of non‐native predators. For example, native prey can fail to recognize and avoid non‐native predators due to a lack of co‐evolutionary history and cue dissimilarity with native predators. This might result in a competitive advantage for non‐native predators. Numerous lady beetle species were globally redistributed as biological control agents against aphids, resulting in novel predator–prey interactions. Here, we investigated the strength of avoidance behavior of the pea aphid (Acyrthosiphon pisum) toward chemical cues of native lady beetles and non‐native Asian Harmonia axyridis and European Coccinella septempunctata and Hippodamia variegata in North America, hypothesizing that cues of non‐native lady beetles induce weaker avoidance behavior than cues of co‐evolved native lady beetles. Additionally, we compared aphid consumption of lady beetles, examining potential predation advantages of non‐native lady beetles. Finally, we compared cue avoidance behavior between North American and European pea aphid populations and aphid consumption of native and non‐native lady beetles in North America and Europe. In North America, pea aphids avoided chemical cues of all ladybeetle species tested, regardless of their origin. In contrast to pea aphids in North America, European pea aphids did not avoid cues of the non‐native H. axyridis. The non‐native H. axyridis and C. septempunctata were among the largest and most voracious lady beetle species tested, on both continents. Consequently, in North America non‐native lady beetle species might have a competitive advantage on shared food resources due to their relatively large body size, compared to several native American lady beetle species. In Europe, however, non‐native H. axyridis might benefit from missing aphid cue avoidance as well as a large body size. The co‐evolutionary time gap between the European and North American invasion of H. axyridis likely explains the intercontinental differences in cue avoidance behavior and might indicate evolution in aphids toward non‐native predators.     

Novel long non‐coding RNA CYB561‐5 promotes aerobic glycolysis and tumorigenesis by interacting with basigin in non‐small cell lung cancer

Abnormally expressed long non‐coding RNAs (lncRNAs) have been recognized as potential diagnostic biomarkers or therapeutic targets in non‐small cell lung cancer (NSCLC). The role of the novel lnc‐CYB561‐5 in NSCLC and its specific biological activity remain unknown. In this study, lncRNAs highly expressed in NSCLC tissue samples compared with paired adjacent normal tissue samples and atypical adenomatous hyperplasia were identified by RNA‐seq analysis. Lnc‐CYB561‐5 is highly expressed in human NSCLC and is associated with a poor prognosis in lung adenocarcinoma. In vivo, downregulation of lnc‐CYB561‐5 significantly decreases tumour growth and metastasis. In vitro, lnc‐CYB561‐5 knockdown treatment inhibits cell migration, invasion and proliferation ability, as well as glycolysis rates. In addition, RNA pulldown and RNA immunoprecipitation (RIP) assays show that basigin (Bsg) protein interacts with lnc‐CYB561‐5. Overall, this study demonstrates that lnc‐CYB561‐5 is an oncogene in NSCLC, which is involved in the regulation of cell proliferation and metastasis. Lnc‐CYB561‐5 interacts with Bsg to promote the expression of Hk2 and Pfk1 and further lead to metabolic reprogramming of NSCLC cells.     

Non‐canonical autophagy functions of ATG16L1 in epithelial cells limit lethal infection by influenza A virus

Influenza A virus (IAV) and SARS‐CoV‐2 (COVID‐19) cause pandemic infections where cytokine storm syndrome and lung inflammation lead to high mortality. Given the high social and economic cost of respiratory viruses, there is an urgent need to understand how the airways defend against virus infection. Here we use mice lacking the WD and linker domains of ATG16L1 to demonstrate that ATG16L1‐dependent targeting of LC3 to single‐membrane, non‐autophagosome compartments – referred to as non‐canonical autophagy – protects mice from lethal IAV infection. Mice with systemic loss of non‐canonical autophagy are exquisitely sensitive to low‐pathogenicity IAV where extensive viral replication throughout the lungs, coupled with cytokine amplification mediated by plasmacytoid dendritic cells, leads to fulminant pneumonia, lung inflammation and high mortality. IAV was controlled within epithelial barriers where non‐canonical autophagy reduced IAV fusion with endosomes and activation of interferon signalling. Conditional mouse models and ex vivo analysis showed that protection against IAV infection of lung was independent of phagocytes and other leucocytes. This establishes non‐canonical autophagy in airway epithelial cells as a novel innate defence that restricts IAV infection and lethal inflammation at respiratory surfaces.     

Individual variation in feeding morphology,not diet,can facilitate the success of generalist species in urban ecosystems

Generalist species dominate urban ecosystems. The success of urban generalists is often related to a plastic diet and feeding traits that allow them to take advantage of a variety of food resources provided by humans in cities. The classification of a species as a generalist is commonly based on mean estimates of diet‐ and feeding‐related traits. However, there is increasing evidence that a generalist population can consist of individual specialists. In such cases, estimates based on mean can hide important individual variation that can explain trophic ecology and the success of urban dwellers. Here, we focus on guppies, Poecilia reticulata, a widespread alien fish species which has invaded both urban and non‐urban systems, to explore the effect of urbanization on individual diet and feeding morphology (cranium shape). Our results show that guppies in urban and non‐urban populations are not individual specialists, having a similar generalist diet despite the high population density. However, there is important individual variation in cranium shape which allow urban guppies to feed more efficiently on highly nutritious food. Our data suggest that individual variation in feeding efficiency can be a critical overlooked trait that facilitates the success of urban generalists.     

Drones,automatic counting tools,and artificial neural networks in wildlife population censusing

1. The use of a drone to count the flock sizes of 33 species of waterbirds during the breeding and non‐breeding periods was investigated.
2. In 96% of 343 cases, drone counting was successful. 18.8% of non‐breeding birds and 3.6% of breeding birds exhibited adverse reactions: the former birds were flushed, whereas the latter attempted to attack the drone.
3. The automatic counting of birds was best done with ImageJ/Fiji microbiology software – the average counting rate was 100 birds in 64 s.
4. Machine learning using neural network algorithms proved to be an effective and quick way of counting birds – 100 birds in 7 s. However, the preparation of images and machine learning time is time‐consuming, so this method is recommended only for large data sets and large bird assemblages.
5. The responsible study of wildlife using a drone should only be carried out by persons experienced in the biology and behavior of the target animals.

     

Are native and non‐native pollinator friendly plants equally valuable for native wild bee communities?

Bees rely on floral pollen and nectar for food. Therefore, pollinator friendly plantings are often used to enrich habitats in bee conservation efforts. As part of these plantings, non‐native plants may provide valuable floral resources, but their effects on native bee communities have not been assessed in direct comparison with native pollinator friendly plantings. In this study, we performed a common garden experiment by seeding mixes of 20 native and 20 non‐native pollinator friendly plant species at separate neighboring plots at three sites in Maryland, USA, and recorded flower visitors for 2 years. A total of 3,744 bees (120 species) were collected. Bee abundance and species richness were either similar across plant types (midseason and for abundance also late season) or lower at native than at non‐native plots (early season and for richness also late season). The overall bee community composition differed significantly between native and non‐native plots, with 11 and 23 bee species being found exclusively at one plot type or the other, respectively. Additionally, some species were more abundant at native plant plots, while others were more abundant at non‐natives. Native plants hosted more specialized plant–bee visitation networks than non‐native plants. Three species out of the five most abundant bee species were more specialized when foraging on native plants than on non‐native plants. Overall, visitation networks were more specialized in the early season than in late seasons. Our findings suggest that non‐native plants can benefit native pollinators, but may alter foraging patterns, bee community assemblage, and bee–plant network structures.     

Psychotherapies for depression: a network meta‐analysis covering efficacy,acceptability and long‐term outcomes of all main treatment types

The effects of psychotherapies for depression have been examined in several hundreds of randomized trials, but no recent network meta‐analysis (NMA) has integrated the results of these studies. We conducted an NMA of trials comparing cognitive behavioural, interpersonal, psychodynamic, problem‐solving, behavioural activation, life‐review and “third wave” therapies and non‐directive supportive counseling with each other and with care‐as‐usual, waiting list and pill placebo control conditions. Response (50% reduction in symptoms) was the primary outcome, but we also assessed remission, standardized mean difference, and acceptability (all‐cause dropout rate). Random‐effects pairwise and network meta‐analyses were conducted on 331 randomized trials with 34,285 patients. All therapies were more efficacious than care‐as‐usual and waiting list control conditions, and all therapies – except non‐directive supportive counseling and psychodynamic therapy – were more efficacious than pill placebo. Standardized mean differences compared with care‐as‐usual ranged from –0.81 for life‐review therapy to –0.32 for non‐directive supportive counseling. Individual psychotherapies did not differ significantly from each other, with the only exception of non‐directive supportive counseling, which was less efficacious than all other therapies. The results were similar when only studies with low risk of bias were included. Most therapies still had significant effects at 12‐month follow‐up compared to care‐as‐usual, and problem‐solving therapy was found to have a somewhat higher long‐term efficacy than some other therapies. No consistent differences in acceptability were found. Our conclusion is that the most important types of psychotherapy are efficacious and acceptable in the acute treatment of adult depression, with few significant differences between them. Patient preference and availability of each treatment type may play a larger role in the choice between types of psychotherapy, although it is possible that a more detailed characterization of patients with a diagnosis of depression may lead to a more precise matching between individual patients and individual psychotherapies.     

Disturbance and the (surprising?) role of ecosystem engineering in explaining spatial patterns of non‐native plant establishment

Different conceptions of disturbance differ in the degree to which they appeal to mechanisms that are general and equivalent, or species‐, functional group‐, or interaction‐specific. Some concepts of disturbance, for example, predict that soil disturbances and herbivory have identical impacts on species richness via identical mechanisms (reduction in biomass and in competition). An alternative hypothesis is that the specific traits of disturbance agents (small mammals) and plants differentially affect the richness or abundance of different plant groups. We tested these hypotheses on a degu (Octodon degus) colony in central Chile. We ask whether native and non‐native forbs respond differently to degu bioturbation on runways versus herbivory on grazing lawns. We ask whether this can explain the increase in non‐native plants on degu colonies. We found that biopedturbation did not explain the locations of non‐native plants. We did not find direct evidence of grazing increasing non‐native herbs either, but a grazing effect appears to be mediated by grass, which is the dominant cover. Further, we provide supplementary evidence to support our interpretation that a key mechanism of non‐native spread is the formation of dry soil conditions on grazing lawns. Thus, ecosystem engineering (alteration of soil qualities) may be an outcome of disturbances, in which each interacts with specific plant traits, to create the observed pattern of non‐native spread in the colony. Based on these results, we propose to extend Jentsch and White (Ecology, 100, 2019, e02734) concept of combined pulse/ disturbance events to the long‐term process duality of ecosystem engineering/ disturbance.     

Lung adenocarcinoma with an uncommon CCDC85A‐ALK fusion responding to alectinib: A case report

A 55‐year‐old Chinese man with a right lung mass and lymph node metastasis (T4N3M0 IIIB) was diagnosed with lung adenocarcinoma after a CT‐guided biopsy. With the wide application of next‐generation sequencing (NGS) in tumour detection, we found a rare CCDC85A‐ALK fusion. The patient received alectinib, which had marked efficacy. This is the first report of a lung adenocarcinoma patient harbouring a new uncommon anaplastic lymphocyte kinase fusion that showed a remarkable response to alectinib. NGS aids in selecting treatment in non‐small cell lung cancer patients.     

Diet overlap among non‐native trout species and native cutthroat Trout (Oncorhynchus clarkii) in two U.S. ecoregions

The invasion of freshwater ecosystems by non‐native species can constitute a significant threat to native species and ecosystem health. Non‐native trouts have long been stocked in areas where native trouts occur and have negatively impacted native trouts through predation, competition, and hybridization. This study encompassed two seasons of sampling efforts across two ecoregions of the western United States: The Great Basin in summer 2016 and the Yellowstone River Basin in summer 2017. We found significant dietary overlaps among native and non‐native trouts within the Great Basin and Yellowstone River Basin ecoregions. Three orders of invertebrates (Ephemeroptera, Trichoptera, and Diptera) composed the majority of stomach contents and were responsible for driving the observed patterns. Great Basin trout had higher body conditions (k), and non‐native Great Basin trout had higher gut fullness values than Yellowstone River Basin trout, indicating a possible limitation of food in the Yellowstone River Basin. Native fishes were the least abundant and had the lowest body condition in each ecoregion. These findings may indicate a negative impact on native trouts by non‐native trouts. We recommend additional monitoring of native and non‐native trout diets, regular invertebrate surveys to identify the availability of diet items, and reconsidering stocking efforts that can result in overlap of non‐native fishes with native cutthroat trout.     

Uniform binding and negative catalysis at the origin of enzymes

Enzymes are well known for their catalytic abilities, some even reaching “catalytic perfection” in the sense that the reaction they catalyze has reached the physical bound of the diffusion rate. However, our growing understanding of enzyme superfamilies has revealed that only some share a catalytic chemistry while others share a substrate‐handle binding motif, for example, for a particular phosphate group. This suggests that some families emerged through a “substrate‐handle‐binding‐first” mechanism (“binding‐first” for brevity) instead of “chemistry‐first” and we are, therefore, left to wonder what the role of non‐catalytic binders might have been during enzyme evolution. In the last of their eight seminal, back‐to‐back articles from 1976, John Albery and Jeremy Knowles addressed the question of enzyme evolution by arguing that the simplest mode of enzyme evolution is what they defined as “uniform binding” (parallel stabilization of all enzyme‐bound states to the same degree). Indeed, we show that a uniform‐binding proto‐catalyst can accelerate a reaction, but only when catalysis is already present, that is, when the transition state is already stabilized to some degree. Thus, we sought an alternative explanation for the cases where substrate‐handle‐binding preceded any involvement of a catalyst. We find that evolutionary starting points that exhibit negative catalysis can redirect the reaction''s course to a preferred product without need for rate acceleration or product release; that is, if they do not stabilize, or even destabilize, the transition state corresponding to an undesired product. Such a mechanism might explain the emergence of “binding‐first” enzyme families like the aldolase superfamily.     

Adverse childhood experiences and mental health problems in a nationally representative study of heterosexual,homosexual and bisexual Danes

Non‐heterosexual persons more often report adverse childhood experiences (ACEs) than heterosexuals, and they generally bear a greater burden of mental health challenges. However, population‐based data on this topic are scarce. In a nationally representative study within the Project SEXUS, one of the world''s largest cohort studies on sexual health, we used data from 57,479 individuals in Denmark to explore the interplay between ACEs and mental health problems among self‐identified heterosexual, homosexual and bisexual persons, and among self‐identified heterosexuals with or without same‐sex sexual experience. Compared to heterosexuals, non‐heterosexual persons were more likely to report most of the studied ACEs, with odds ratios (ORs) for the ACE category “abuse” ranging from 1.38 to 1.75 for homosexual women, from 1.76 to 2.65 for homosexual men, from 2.52 to 3.64 for bisexual women, and from 1.58 to 6.07 for bisexual men. Furthermore, non‐heterosexual persons had consistently and statistically significantly higher odds for mental health problems (ORs: 1.50 to 4.63). Combinations of ACEs with a non‐heterosexual identity resulted in markedly elevated odds for mental health problems, particularly among bisexual individuals. This included high odds for suicidal thoughts/attempts among bisexual persons with a history of “neglect” (women: OR=12.82; men: OR=35.24) and “abuse” (women: OR=11.81; men: OR=11.65). Among self‐identified heterosexuals, combinations of ACEs with same‐sex sexual experience were associated with consistently elevated odds for mental health problems (ORs: 2.22 to 12.04). The greater burden of ACEs among self‐identified homosexuals and, most notably, bisexuals may account for part of their excess risk of mental health problems. These findings emphasize the public health importance of preventive measures to minimize the burden of ACEs and avert their harmful long‐term effects. Moreover, they highlight the need to safeguard the welfare of children and adolescents with non‐conforming expressions of sexuality.     

Nicking and fragmentation are responsible for α‐lactalbumin amyloid fibril formation at acidic pH and elevated temperature

Amyloid fibrils are ordered aggregates that may be formed from disordered, partially unfolded, and fragments of proteins and peptides. There are several diseases, which are due to the formation and deposition of insoluble β‐sheet protein aggregates in various tissue, collectively known as amyloidosis. Here, we have used bovine α‐lactalbumin as a model protein to understand the mechanism of amyloid fibril formation at pH 1.6 and 65°C under non‐reducing conditions. Amyloid fibril formation is confirmed by Thioflavin T fluorescence and atomic force microscopy (AFM). Our finding demonstrates that hydrolysis of peptide bonds occurs under these conditions, which results in nicking and fragmentation. The nicking and fragmentation have been confirmed on non‐reducing and reducing gel. We have identified the fragments by matrix‐assisted laser desorption ionization‐time of flight (MALDI‐TOF) mass spectrometry. The fragmentation may initiate nucleation as it coincides with AFM images. Conformational changes associated with monomer resulting in fibrillation are shown by circular dichroism and Raman spectroscopy. The current study highlights the importance of nicking and fragmentation in amyloid fibril formation, which may help understand the role of acidic pH and proteolysis under in vivo conditions in the initiation of amyloid fibril formation.