Category: 3. Business

It’s become the playbook for big Australian companies that have customer data stolen in a cyber-attack: call in the lawyers and get a court to block anyone from accessing it.

Qantas ran it recently after suffering a major cybersecurity attack that accessed the frequent flyer details of 5 million customers.

The airline joined the long list of companies in Australia, dating back to the HWL Ebsworth breach in 2023, to go to the NSW supreme court to obtain an injunction against “persons unknown” – banning the hackers (and anyone else) from accessing or using the data under threat of prosecution.

Of course, it didn’t stop hackers leaking the customer data on the dark web a few months later.

But it might have come as a surprise when ID protection company Equifax this month began alerting Qantas customers that their data had been leaked – since access to the data was supposedly banned.

This highlights the major flaw in the injunction scheme. Qantas argues the injunction protects customers, but cybersecurity experts warn that in practice it has the opposite effect: scammers will ignore it, while organisations based in Australia and operating within the law will not be able to verify the data and report on it.

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Troy Hunt, an Australian who operates the HaveIBeenPwned website which notifies users when their information appeared in breaches, is frustrated that he has not been able to include the breach in his searchable database.

“Clearly the injunction has not stopped even legally operating organisations from accessing the data and communicating with the customers,” he said.

“[Qantas is] obviously trying to minimise damage, and they will inevitably get raked over the coals with class actions, because it happens to every big company that has a breach now … but there is just no measurable, practical benefit that anyone can assign to keeping this data out of the hands of people like [me], whilst it’s in the hands of people who are now abusing it.”

Hunt noted the irony that Qantas’s cybersecurity incident statement on its website links out to government resources for customers caught up in a breach. Those resources advise customers to visit Hunt’s website so they can better protect themselves by being aware of what information is out there.

How Equifax approached the injunction is unclear. The company said it uses the cybersecurity company Norton to monitor the dark web. Norton’s parent company Gen Digital is based in the US and Czechia while Equifax is US-based.

Norton did not deny it had accessed the data when asked twice by Guardian Australia, saying in a statement it is “contractually obligated to notify customers” when their information is posted on the dark web.

“These alerts are part of our ongoing commitment to help victims of a data breach protect their personal information and respond quickly if their data is at risk,” the spokesperson said. “This service operates under strict business, privacy, and compliance standards to ensure accuracy and lawful handling of all data sources.”

Qantas would not confirm if it was considering pursuing companies for potential contraventions of the injunction, but indicated it was monitoring third-parties and would consider them on a case-by-case basis.

“We are aware of notifications being sent to some of our customers by a third-party providers. These notifications include types of personal information that was not held in the system impacted in our July cyber incident,” the spokesperson said.

According to screenshots from the Telegram group run by the hackers, posted this month by Hunt, the hackers are aware of the limitations of the injunction.

“qantas why are you lying to your citizens?” the message states. “all your injunction does is prevent media/journalists.”

“YOUR data WILL be released and it WILL BE accessed.”

In an interview with Bloomberg TV’s Wall Street Week on Friday, he said the obvious way to make money off AI investments, aside from charging fees to use chatbots, is to replace workers with something cheaper.

Hinton, whose work has earned him a Nobel Prize and the moniker “godfather of AI,” added that while some economists point out previous disruptive technologies created as well as destroyed jobs, it’s not clear to him that AI will do the same.

“I think the big companies are betting on it causing massive job replacement by AI, because that’s where the big money is going to be,” he warned.

Just four so-called AI hyperscalers—Microsoft, Meta, Alphabet and Amazon—are expected to boost capital expenditures to $420 billion next fiscal year from $360 billion this year, according to Bloomberg.

Meanwhile, OpenAI alone has announced a total of $1 trillion in infrastructure deals in recent weeks with AI-ecosystem companies like Nvidia, Broadcom and Oracle.

When asked if such investments can pay off without destroying jobs, Hinton replied, “I believe that it can’t. I believe that to make money you’re going to have to replace human labor.”

The remarks echo what he said in September, when he told the Financial Times that AI will “create massive unemployment and a huge rise in profits,” attributing it to the capitalist system.

In fact, evidence is mounting that AI is shrinking opportunities, especially at the entry level, and an analysis of job openings since OpenAI launched ChatGPT shows they plummeted roughly 30%.

And this past week, Amazon announced 14,000 layoffs, largely in middle management. While CEO Andy Jassy said the decision was due to “culture” and not AI, a memo he sent in June predicted a smaller corporate workforce “as we get efficiency gains from using AI extensively across the company.”

Despite the potential downside for workers, Hinton also sees benefits from AI. When asked if he would go back in time and stop AI from developing, he paused and said he doesn’t know.

“It’s not like nuclear weapons, which are only good for bad things,” he explained. “It’s a difficult decision because it can do tremendous good in healthcare and education. It’ll do tremendous good, and in fact if you think about it increasing productivity in many, many industries, that should be good.”

The problem ultimately is not due to AI itself, but “on how we organize society,” Hinton added.

PERK promotes myogenesis in myoblasts subjected to physiological levels of ER stressors

To investigate the role of adaptive UPR^ER during myogenesis, C2C12 myoblasts were exposed to a range of concentrations of the ER stressor tunicamycin (TM) for 8 h, followed by differentiation into myotubes (Supplementary Fig. 1A). Notably, treatment of myoblasts with 0.2 μg/ml TM significantly enhanced the myogenic potential of myoblasts. However, concentrations above 0.4 μg/ml were detrimental and promoted cell death (Supplementary Fig. 2A). To determine if the response was an ER stress response or specific to TM, the SERCA inhibitor thapsigargin (TG) was also used. A low concentration of TG (10 nM) also promoted an increase in myogenic parameters (Supplementary Fig. 2B). Inhibition of PERK with AMG PERK44 (AMG) resulted in no difference in myogenesis parameters compared to controls (Fig. 1A). Furthermore, the combined AMG + TM treatment group (AMG + TM) exhibited no change when compared to the AMG group (Fig. 1A). However, inhibition of IRE1α with MKC8866 (MKC) led to a significant decrease in all the parameters compared to controls (Fig. 1A). Nevertheless, the MKC + TM group exhibited an increase in myogenic parameters compared to MKC alone (Fig. 1A). Interestingly, TM-treated myoblasts exhibited a significant decrease in cell viability, with an even greater decrease observed in the AMG + TM group (Supplementary Fig. 2C).

PCR analysis confirmed activation of the IRE1α arm of the UPR^ER, with a shift toward the spliced, active form of XBP1 (XBP1s) following TM treatment (Fig. 1B). Western blot analysis of UPR^ER markers revealed no significant changes in the ER chaperone GRP78, though there was an increasing trend in eIF2α phosphorylation (Supplementary Fig. 2D). There was an increase in the levels of ATF4 in MKC + TM-treated cells, and increasing trend in eIF2α phosphorylation, while CHOP increased following TM treatment and was elevated in both AMG + TM and MKC + TM groups (Fig. 1C–E). To evaluate the different UPR pathways, we performed a qPCR analysis of markers of the mammalian UPR^ER (Hspa5, Dnajc3, Ddit3, Herpud1, Pdia4 and Edem1) [44] and UPR^mt (Atf5, Lonp1, Hspe1, Hspa1a, Hspd1 and Hsp90aa1) [45]. A similar response in all UPR^ER markers was observed, a slight increase in gene expression with the TM treatment, however there was a significant increase in levels of UPR^ER genes following TM treatment in combination with the PERK inhibitor (Supplementary Fig. 2E). The expression of UPR^mt genes Atf5 and Lonp1, follow the same pattern as the UPR^ER markers, however, the gene expression of Hspe1, Hspa1a, Hspd1 and Hsp90aa1 did not change (Supplementary Fig. 2F). Similar to inhibition of PERK with AMG44, knock-down of PERK (siPERK) in myoblasts decreased myotube diameter following differentiation. Knock-down of PERK or CHOP combined with the TM treatment, exhibited a reduction in myotube area and fusion index compared to siRNA CNT (Supplementary Fig. 2E). Additionally, TM treatment of differentiated, untreated myotubes did not result in any significant differences in the myotube parameters (Fig. 1F). Collectively, these findings demonstrate that inducing mild ER stress in myoblasts improved myogenesis and identified an essential role of PERK in adaptive UPR signalling.

PERK promotes MERCS assembly and mitochondrial adaptations during adaptive UPR

The effects of TM treatment (0.2 μg/ml for 8 h) and/or PERK inhibition on mitochondrial content and function were evaluated in myoblasts (Fig. 1A). MitoTracker Green staining was used to assess mitochondrial content and increased with TM treatment. No differences were observed between the AMG group and the controls (Fig. 2A). Mitochondrial ROS was assessed using MitoSOX staining and there was no change in MitoSOX staining in the TM and AMG groups. However, an increase in fluorescence was observed in the AMG + TM group (Fig. 2A). Additionally, MitoTracker Green and TMRE staining was used to assess mitochondrial membrane potential (ΔΨm), there was a significant increase in both MitoTracker Green and TMRE staining following TM treatment. The ratio of the intensities indicated that TM treatment promoted mitochondrial biogenesis and enhanced ΔΨm (Supplementary Fig. 2F). Both the mitochondrial fusion protein MFN2, which plays a role in mitochondrial-ER tethering [46, 47], and the mitochondrial content indicator TOM20 increased with TM treatment (Fig. 2B, C). The oxygen consumption rate of C2C12 myoblasts demonstrated enhanced basal and maximal respiration, non-mitochondrial respiration, and ATP production in TM-treated cells compared to controls, AMG, and AMG + TM groups (Fig. 2D, E). Furthermore, transmission electron microscopy (TEM) organelle ultrastructure analysis of myoblasts demonstrated an increase in the membrane surface area of mitochondria (S_A), mitochondrial elongation (aspect ratio) following TM treatment; these adaptations were blocked by PERK inhibition (Fig. 2F, G). Notably, these mitochondrial adaptations were preserved following differentiation into myotubes (Fig. 2I, J). Additional parameters, including mitochondrial volume fraction, cristae membrane density (S_A IMM/OMM) and ER surface area, increased in both myoblasts and subsequent myotubes following TM treatment (Supplementary Fig. 2G). The evaluation of MERCS surface area demonstrated an expansion of mitochondrial and ER membranes that are in close proximity following TM treatment. The formation of these expanded subdomains was blocked by inhibition of PERK (Fig. 2F–H). The increase in MERCS surface area was sustained after differentiation into myotubes (Fig. 2I–K). Together, the data demonstrates that PERK is required during adaptive UPR^ER to form MERCS and increase mitochondrial content and function in myoblasts, which were maintained following differentiation into myotubes.

PERK is required for adaptive UPR^ER activation of autophagy and mitophagy

To determine the mechanisms underlying the adaptations promoted by the UPR^ER in myoblasts, proteomic changes induced by an 8 h treatment with 0.2 μg/ml TM was performed. Global label-free proteomics quantified 2513 proteins, 71 proteins were significantly upregulated and 27 downregulated following TM treatment (Fig. 3A). Enrichment analysis of the 71 significantly upregulated proteins identified several pathways and proteins involved in Mitophagy and mitochondrial ADP/ATP transport (ADRO, TPP1, TXTP, MTDC, SYHM, RM12, KAPCA, ADT1 and ADT2), indicating active remodelling of the mitochondrial network and mitochondrial respiration (Supplementary Fig. 2J, K). Also, upregulated were proteins related to Cell cycle and differentiation (PTBP3, RCC1, CCNK, MD1L1 and MYH10) (Fig. 3A, B). One of the most significantly upregulated proteins was ERD22, a member of the KDELR family highly involved in the maintenance of ER homeostasis during adaptive UPR^ER signalling [48]. There was a significant increase in gene expression of Kdelr2 following TM treatment (Supplementary Fig. 2L). No significant pathway enrichment was identified among the 27 downregulated proteins, which spanned diverse biological processes, including mitochondrial RNA processing and protein translation (4EBP1, MRT4, RPR1B and EI2BA) (Fig. 3A).

To further explore mitochondrial turnover, live-cell imaging was performed using the mitophagy reporter plasmid [Cox8-EGFP-mCherry] [49], transfected into myoblasts before TM treatment. The reporter contains a pH-sensitive GFP and a pH-stable RFP to track mitochondrial turnover; GFP fluorescence is quenched in acidic lysosomes, while RFP remains stable, allowing an estimation of mitophagy (Fig. 3C). TM treatment significantly reduced the GFP/RFP ratio compared to the control group, indicating an induction of mitophagy (Fig. 3D, E). Western blot analysis of autophagy-related proteins revealed that ULK1 levels were elevated in the TM group compared to the CNT, AMG, and AMG + TM groups (Supplementary Fig. 2M). p62 levels increased in the AMG and AMG + TM groups compared to the CNT and TM groups, indicating reduced autophagic flux (Fig. 3F). Finally, increased expression of ATG5 (Supplementary Fig. 2N) and LC3-II/I (Fig. 3G) in the TM group indicates enhanced autophagosome formation. Ultrastructure analysis of the autophagic bodies by TEM, evaluated changes in the volume fraction of lysosomes, autophagosomes and autolysosomes from myoblasts and myotubes. Myoblasts exhibited an apparent increase in the volume fraction of lysosomes and autolysosomes in the TM group compared to CNT, AMG and AMG + TM groups (Fig. 3H). In contrast, myotube analysis indicated no change in lysosome or autolysosomes volume fractions between groups, although there was a decrease in autophagosome volume in the TM group compared to the CNT and AMG + TM groups (Fig. 3I). Changes in [Ca²⁺]_i showed a slight, but not significant, increase in the maximal increase of store-operated calcium entry (SOCE) from the TM-treated myoblasts compared to controls (Supplementary Fig. 2O). Additionally, ER volume was assessed using an ER-targeting plasmid [ER (KDEL)-mNeonGreen] to estimate the ER membrane area relative to cellular surface area, revealing a significant expansion in ER membrane area following TM treatment (Supplementary Fig. 2P). Overall, proteomic and ultrastructural analyses revealed that TM treatment induces mitophagy and enhances autophagic flux during myogenesis. These adaptations depend on PERK activity and potentially play a crucial role in regulating organelle turnover.

Early-life induction of the UPR^ER extends lifespan and preserves healthspan in C. elegans

C. elegans was used as a physiological model to assess the role of UPR^ER signalling in a whole organism. To replicate the in vitro myogenesis model, worms were treated at an early developmental stage. Following bleaching, embryos were harvested and exposed to 1.25 μg/ml TM in S Medium for 24 h and larvae plated and allowed to develop normally (Supplementary Fig. 1B) [50]. There was an increase in the longevity of TM-treated worms compared to controls (Fig. 4A). To determine if the response was an ER stress response or specific to TM, the SERCA inhibitor TG was used. Treatment with 150 nM of TG also promoted an increase in lifespan and increased filamentous mitochondrial network (Supplementary Fig. 3A, B). Reproductive potential was significantly reduced in the TM group (Fig. 4B). Importantly, this decrease in progeny was not associated with increased embryo lethality (Supplementary Fig. 3C). Interestingly, TM-treated worms exhibited increased body length compared to controls (Fig. 4C). The activation of both the UPR^ER and the UPR^mt was confirmed using transcriptional GFP reporter strains for hsp-4 (orthologue of mammalian HSPA5/Grp78) and hsp-6p (ortholog of the human mtHSP70). TM treatment resulted in greater UPR^ER and UPR^mt activation capacity compared to controls (Fig. 4D, E). Analysis of the IRE-1 branch of the UPR revealed increased splicing of XBP-1 in TM-treated worms (Fig. 4F). The survival of the worms against a range of different toxicants was assessed. The resistance of the TM treated group against sodium arsenite, the redox cycler Paraquat and the organic peroxide tBuOOH was higher than the control group at adult day 1 (Fig. 4G–I). CeleST (C. elegans Swim Test) was performed to assess physical fitness and mobility during ageing [42]. The fitness levels were evaluated on days 1, 5, 10 and 15 of adulthood. Fitness markers including wave initiation rate, travel speed and activity index decreased with age in both groups but remained significantly higher in TM-treated worms compared to controls at all time points (Fig. 4J, Supplementary Fig. 3D, E). Conversely, frailty markers including stretch, average body curvature and curling increased with age in both groups, but these were consistently lower in the TM group, indicating better maintenance of physical health in the TM-treated groups (Fig. 4K, Supplementary Fig. 3F, G). The results demonstrate that early-life UPR^ER activation extends lifespan, increases stress resistance and promotes healthspan by preserving physiological fitness and reducing frailty during ageing.

Early-life induction of the UPR promotes lifespan extension in C. elegans through modulation of lipid metabolism and lysosome activation

As TM treatment of myoblasts resulted in improved myogenesis, while treatment of myotubes was detrimental, we assessed whether the adaptations promoted by the UPR^ER during early nematode development were similar to TM treatment of adult worms. TM treatment (1.25 and 5 μg/ml for 24 h) was performed during L4 to adult Day1 developmental stage of C. elegans. Both TM treatments (1.25 and 5 μg/ml) at L4 stage resulted in a significant reduction in lifespan (Fig. 5A), along with decreased resistance to paraquat and sodium arsenite (Fig. 5B, C). Interestingly, no change in resistance to sodium arsenite was observed in worms treated with the lower TM dose (1.25 μg/ml) (Fig. 5B). This highlights that the beneficial effects of TM treatment occurred only at an early developmental stage and treatment of adult worms resulted in negative effects similar to the myogenesis model.

RNA sequencing was performed to determine the molecular mechanisms underlying the different responses dependent on the developmental stage. A transcriptome analysis was conducted on Day 1 adults that were treated with TM 1.25 μg/ml at embryonic stage (EMBRYO group) (Fig. 5D) or treatment with TM at L4 stage (L4 group) (Fig. 5E). Differentially expressed genes (DEGs) were analysed (Log₂-fold change >1 and adjusted p-value < 0.05) from EMBRYO (FDR = 2.38%) and L4 separately (FDR = 1.1%). DEGs from EMBRYO-treated groups reported 1249 upregulated and 2042 downregulated genes. Functional enrichment analysis of the upregulated genes identified pathways related to lipid metabolism and lysosome activation (Fig. 5F). Both pathways have previously been described as key intermediates in the UPR regulation of ageing [51]. Gene set enrichment analysis (GSEA) indicated enhanced expression of genes associated with autophagy (lysosomal pathways) and mitochondrial activity (oxidative phosphorylation) (Supplementary Fig. 3I). Lipid biosynthesis emerged as the most prominent upregulated pathway and there was an increase in most intermediates of this pathway (Supplementary Fig. 4A). Gene set enrichment analysis (GSEA) indicated positive enrichment of genes associated with autophagy (lysosomal pathways) and mitochondrial activity (oxidative phosphorylation) (Supplementary Fig. 4B). In contrast, the L4 group resulted in 146 upregulated and 811 downregulated genes (Fig. 5F). Functional enrichment of the downregulated genes highlighted pathways involved in lipid metabolism and C. elegans longevity (Fig. 5G). Pathway enrichment analysis identified a reduction in the expression of intermediates in the lipid biosynthesis pathway in the L4 treated group (Supplementary Fig. 4D). GSEA revealed decreased expression of genes related to autophagy (lysosomes) and mitophagy in L4-treated worms (Supplementary Fig. 4E). These results were validated by qPCR of genes involved in lysosomal function (vha-6, asah-1 and cpr-1) and lipid metabolism (acox-1.5, elo-2 and fat-5), increased expression in the EMBRYO treated group and decreased expression of fat-5 and vha-6 in the L4 treated groups (Supplementary Fig. 4G, H). Finally, comparing the functional enrichment of downregulated genes in the EMBRYO group with the upregulated genes in the L4 group demonstrated shared pathways related to metabolism (Supplementary Fig. 3C–F). These findings suggest that UPR^ER activation during early development promotes beneficial adaptations that contribute to lifespan extension, whereas induction later in life leads to a decline in similar protective mechanisms, particularly affecting lipid metabolism and autophagy.

RNA sequencing highlighted the effects of TM treatment on lipid metabolism and the lysosome. Oil Red O staining, which stains neutral triglycerides and lipids, was performed in the different groups [40]. Interestingly, TM treatment decreased lipid staining in the EMBRYO group compared to the controls (Fig. 5J). However, in the L4 group, TM treatment increased lipid staining compared to the control group (Fig. 5J), supporting the transcriptomic data of increased expression of lipid-related genes in embryo-treated worms but a decrease in L4 treated worms. The lysosomal content and acidity were estimated using the LysoSensor green/LysoTracker red (LSG/LSR) ratio in the intestine [52]. Results demonstrated decreased pH in the lysosomes of N2 worms subjected to the TM treatment at the embryonic stage. No changes in lysosomal pH were reported in the animals treated at the L4 stage (Fig. 5K). An upregulation of lysosomal genes and increased intestinal lysosomal acidity in response to UPR^ER activation has previously been demonstrated to improve proteostasis and longevity in C. elegans [53]. RNAi knockdown of lmp-1 (ortholog of LAMP1, a lysosomal membrane protein important for lysosomal function) had no effect on longevity or in combination with TM treatment (Supplementary Fig. 3L). RNAi knockdown of lmp-1 also resulted in a more fragmented mitochondrial network after TM treatment (Supplementary Fig. 3M). These results suggest activation of autophagy is required for the adaptive UPR^ER increase in longevity and mitochondrial morphology. Overall, the effects of TM treatment were dependent on the developmental stage and resulted in the opposite expression of genes involved in lipid metabolism, and subsequent effects on longevity and oxidative stress resistance. The results support the findings in the myogenesis model and highlight increased plasticity for adaptations following UPR^ER induction at early developmental stages.

Early-life UPR induction stimulates MERCS formation, mitochondrial capacity and mitophagy in C. elegans

In order to determine if there was a similar adaptive response in nematodes to the myogenesis model following TM treatment, the effects on mitochondrial content were assessed using a transcriptional Pcox-4::GFP reporter [54]. The fluorescence intensity increased with TM treatment (Fig. 6A). MitoTracker Red staining was used to estimate mitochondrial membrane potential and there was a significant increase in the fluorescence intensity following treatment with TM (Fig. 6B), which was also observed using TMRM staining (Supplementary Fig. 3N). In addition, mitochondrial superoxide production was evaluated by MitoSOX staining, demonstrated a reduction in the TM group (Supplementary Fig. 3O). Mitochondrial respiration was assessed using adult day 1 C. elegans following treatment with TM at the embryo stage. TM-treated worms showed a slight, non-significant increase in both basal and maximal OCR, and there was a significant increase in non-mitochondrial respiration (Fig. 6C). TEM was used to analyse the mitochondrial ultrastructure of body wall muscle cells from C. elegans (Supplementary Fig. 1D). Stereological analysis revealed increased mitochondrial surface area in the TM group (Fig. 6D). Moreover, mitochondria from the TM treatment group, exhibited an increased aspect ratio, indicating more elongated, fused mitochondria. The surface area ratio between the inner (IMM) and outer mitochondrial membranes (OMM) was also significantly increased, reflecting a denser IMM structure (Fig. 6D), which may contribute to the observed improvements in mitochondrial function.

Mitochondrial turnover was assessed using the pmyo-3::TOMM-20::Rosella reporter strain, containing a pH-sensitive Rosella biosensor composed of a pH-sensitive GFP and a pH-stable RFP to track mitochondrial turnover in body wall muscle [55]. TM-treated worms exhibited a significantly lower GFP/DsRed fluorescence ratio, indicating increased mitophagy and enhanced mitochondrial turnover during adulthood (Fig. 6E). RNAi knockdown of pdr-1 (ortholog of Parkin, regulator of mitochondrial turnover) had no effect on longevity but in combination with TM treatment decreased lifespan compared with TM alone, RNAi knockdown of pdr-1 resulted in a more fragmented mitochondrial network (Supplementary Fig. 3J, K). RNAi knockdown of lgg-1 (ortholog of LC3, required for autophagosome maturation) following TM treatment resulted in no change in lifespan or on mitochondrial morphology (Supplementary Fig. 3H, I). These results indicate activation of mitophagy is required for the increased longevity and changes in mitochondrial morphology following adaptive UPR^ER signalling.

As MERCS are essential for mitochondrial fission, TEM analysis was performed to quantify the surface area of mitochondria in close contact with the ER (<50 nm). TM-treated worms resulted in a marked increase in MERCS surface area compared to controls (Fig. 6F). Mitochondrial morphology was evaluated during ageing using a genetic pmyo-3::mitoGFP reporter, for visualisation of the mitochondrial network from body wall muscle. Images were taken on days 2, 4 and 6 of adulthood and classified as punctate, intermediate or filamentous, depending on the mitochondrial network morphology [56]. Ageing resulted in a progressive shift towards more punctate (fragmented) networks (Fig. 6G). In contrast, TM-treated worms demonstrated a delayed onset of mitochondrial fragmentation, maintaining more filamentous and intermediate networks at all time points assessed (Fig. 6G). In order to investigate whether this response is redox dependent, the antioxidant N-Acetyl Cysteine (NAC) was used. TM treatment during early development in combination with NAC resulted in the repression of lifespan extension and increased mitochondrial fragmentation, indicating the beneficial adaptive UPR response has a redox component (Supplementary Fig. 3P, Q). Together, the data demonstrate that, similar to myoblasts, early-life TM treatment helps preserve mitochondrial network integrity and morphology during ageing, potentially through the formation of MERCS and regulation of mitochondrial turnover.

PEK-1 (UPR^ER) and ATFS-1 (UPR^mt) crosstalk regulates the adaptations promoted by the UPR in C. elegans

To elucidate the mechanisms underlying UPR-mediated mitochondrial and physiological adaptations, we used mutant strains for the UPR^ER and UPR^mt signalling arms to evaluate their response to early-life TM treatment. Both the ire-1(v33) and xbp-1(tm2482) mutant strains were viable. However, TM treatment resulted in developmental arrest at the larval stage (Supplementary Fig. 3A). This phenotype has previously been reported, generation of double mutants of ire-1 or xbp-1 with atf-6 or pek-1 promoted developmental arrest at larvae stage L2 [57]. highlighting the critical role of the IRE-1 arm in the resistance to ER stress. Similar to the N2 strain, treatment of the atf-6 (ok551) mutant strain with TM resulted in increased longevity and stress resistance to sodium arsenite but not paraquat but increased MitoTracker Red staining (Supplementary Fig. 5B). Interestingly, following TM treatment, atf-6 mutants exhibited a reduction in size at day 1 of adulthood, implying delayed development (Supplementary Fig. 5B). As the increased longevity and survival following TM treatment were also observed in N2 strain (Fig. 4A, G, H), it would suggest they are independent of ATF-6 signalling. Following TM treatment of the PEK-1 loss-of-function mutant strain (pek-1 (ok275)), there was a decrease in lifespan (Fig. 7A), resistance to paraquat was reduced but resistance to sodium arsenite was unaltered (Supplementary Fig. 5C). There was a decrease in MitoTracker Red fluorescence intensity in the TM-treated pek-1 mutants (Fig. 7B). The pek-1 mutant strain also had a reduction in size following TM treatment (Supplementary Fig. 5C). TM treatment of pek-1 mutant strain produced an opposite phenotype compared to the N2 strain, highlighting its importance for this adaptive response. TM treatment of ATFS-1 mutant (atfs-1 (tm4525)) also exhibited a decrease in lifespan (Fig. 7C); however, resistance to oxidative stressors was unaltered (Supplementary Fig. 5D). The ΔΨm of the TM-treated atfs-1 mutant strain decreased following TM treatment (Fig. 7D). Interestingly, TM-treated atfs-1 mutant exhibited no differences in size compared to the control group (Supplementary Fig. 5D). The results highlight that TM treatment of the atfs-1 mutant strain had similar results to the pek-1 mutant and an opposite phenotype to N2 WT strain, highlighting its importance for this adaptive response.

A double pek-1;atfs-1 mutant was generated to clarify the possible interaction of these signalling pathways and potential crosstalk of the UPR^ER and UPR^mt. The phenotypic effects of TM treatment in the N2 wild-type strain, pek-1, atfs-1 and pek-1;atfs-1 strains were determined. All mutant strains had decreased lifespan compared to the WT group which had increased longevity (Fig. 7E, F, Supplementary Table 1). Interestingly, TM increased the size of N2 worms, but this effect was reversed in the pek-1 mutant and absent in the atfs-1 mutant (Supplementary Fig. 5E). The pek-1;atfs-1 double mutant exhibited a more severe reduction in body size than either single mutant, both under basal conditions and following TM treatment (Supplementary Fig. 5E). All mutant strains treated with TM had decreased resistance to sodium arsenite and paraquat compared to N2 strain (Supplementary Fig. 5E, Supplementary Tables 2, 3).

Mitochondrial membrane potential was significantly reduced in both the atfs-1 and pek-1;atfs-1 mutants compared to N2, while no differences were observed between the untreated N2 and the pek-1 mutant strain (Fig. 7G). TM treatment only increased mitochondrial membrane potential in N2 WT worms, with all mutants displaying lower MitoTracker Red staining post-TM treatment (Fig. 7G). To assess mitochondrial morphology in body wall muscle, strains were crossed with the pmyo-3::mitoGFP reporter strain and imaged at day 4 of adulthood. Control N2 WT worms had a balanced mitochondrial network with approximately 20% filamentous, 40% intermediate, and 40% punctate mitochondria. The pek-1 and atfs-1 mutants demonstrated increased punctate or more fragmented mitochondrial network (Fig. 7H). However, the pek-1;atfs-1 double mutant displayed a highly disrupted mitochondrial network, with no filamentous mitochondria present. TM treatment increased the percentage of filamentous mitochondria in N2 WT worms (~25%) and reduced the percentage of mitochondrial punctate (~25%). In contrast, the mitochondrial morphology of pek-1 and atfs-1 single mutants remained unchanged following TM treatment, while the pek-1;atfs-1 mutant exhibited further mitochondrial fragmentation (Fig. 7H). Finally, to assess physiological activity, CeLeST analysis was performed. In untreated conditions, fitness-related parameters were similar between WT, pek-1, atfs-1, and pek-1;atfs-1 mutants, while frailty parameters were slightly higher in the mutant strains (Fig. 7I, J, Supplementary Fig. 3F). Following TM treatment, WT worms had increased wave initiation rate, travel speed and activity index indicating increased fitness, this was not observed in any of the mutant strains (Fig. 7, Supplementary Fig. 3F). To dissect the mechanistic basis of this crosstalk we evaluated the cellular response to early-life ER stress. Downstream markers of the UPR^ER and UPR^mt (hsp-4, a canonical UPR^ER target, and hsp-6, a mitochondrial chaperone and target of ATFS-1) exhibited a clear increase in both following TM treatment in the N2 strain. However, mutant strains pek-1(-), atfs-1(-) and pek-1(-);atfs-1(-) did not increase hsp-4 or hsp-6 following TM treatment suggesting both PEK-1 and ATFS-1 are required for adaptive UPR activation (Supplementary Fig. 5G).

Together, these results highlight the critical roles of PEK-1 and ATFS-1 in regulating lifespan, stress resistance and mitochondrial dynamics. The results indicate that under mild ER stress conditions, PEK-1 from the UPR^ER establishes crosstalk with ATFS-1 from the UPR^mt pathway to promote increased lifespan, improved healthspan, enhanced stress resistance and greater mitochondrial function. Moreover, based on the gene expression profile of UPR markers, the slight increase in these markers with the TM treatment elicit an adaptive UPR activation, whereas the more drastic activation of the markers in the AMG + TM group could indicate an induction of maladaptive UPR signalling.

In summary, the data presented using both an in vitro myogenesis model and in vivo whole organism C. elegans, reveals that adaptive UPR^ER signalling through PERK and UPR^mt, induces the assembly of MERCS and regulates mitochondrial adaptations following a low dose of an ER stressor at an early developmental stage. These adaptations promote myogenesis and extension of lifespan in C. elegans by enhancing organelle turnover and mitochondrial function, demonstrating the essential crosstalk between ER and mitochondrial stress responses in maintaining cellular and organismal homeostasis under physiological stress conditions.

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With shares lagging despite solid long-term returns and new initiatives, investors may wonder whether Rush Enterprises is trading at a compelling discount or if the market is already factoring in any rebound in future truck demand.

With Rush Enterprises closing at $49.41 compared to a widely followed narrative fair value of $60.00, the stock stands out for those seeking value among commercial vehicle retailers. The difference between current price and narrative value shines a spotlight on what underpins this estimate.

Extended regulatory and trade policy uncertainty is causing customers to delay new vehicle purchases, leading to aging truck fleets that require increased parts and service work. This supports stable or rising revenue and margins from Rush’s high-margin aftermarket business in the near term, which already accounts for over 60% of gross profit.

Read the complete narrative.

Wondering what drives this bold valuation gap? The secret mix: revenue stream shifts, new margin records, and a forecast that could surprise the entire transportation sector. Dig into the details behind these assumptions and see why the consensus is betting on a future far brighter than the recent past.

Result: Fair Value of $60.00 (UNDERVALUED)

Have a read of the narrative in full and understand what’s behind the forecasts.

However, persistent regulatory uncertainty or a prolonged “freight recession” could strain both truck sales and aftermarket revenue. This would challenge the current optimism in forecasts.

Find out about the key risks to this Rush Enterprises narrative.

If you see the story differently or want to reach your own conclusions, you can quickly build your personalized narrative from the data in just a few minutes. Do it your way

A great starting point for your Rush Enterprises research is our analysis highlighting 2 key rewards and 1 important warning sign that could impact your investment decision.

Take control of your portfolio and gain an edge by researching stocks that align with your goals. The right ideas can make all the difference in seizing opportunities ahead of the crowd.

This article by Simply Wall St is general in nature. We provide commentary based on historical data and analyst forecasts only using an unbiased methodology and our articles are not intended to be financial advice. It does not constitute a recommendation to buy or sell any stock, and does not take account of your objectives, or your financial situation. We aim to bring you long-term focused analysis driven by fundamental data. Note that our analysis may not factor in the latest price-sensitive company announcements or qualitative material. Simply Wall St has no position in any stocks mentioned.

Companies discussed in this article include RUSHA.

Have feedback on this article? Concerned about the content? Get in touch with us directly. Alternatively, email editorial-team@simplywallst.com

Story Continues

The most common refrain so far this corporate earnings season is that the U.S. economy is increasingly divided between wealthy consumers and everyone else.

While many Americans tighten their belts amid rising costs and a weakening job market, a fortunate few are still spending freely, insulated by a financial cushion of stock market gains and years of rising home values.

The split is having an outsize impact on consumer-facing companies, but those effects aren’t being felt evenly across brands.

McDonald’s CEO Chris Kempczinski said the “two-tier economy” was a major factor in the fast-food giant’s decision to revive its “Extra Value Meal” combos last month.

“Traffic for lower-income consumers is down double digits,” Kempczinski told CNBC in September. “We needed to step in.”

Coca-Cola’s chief operating officer, Henrique Braun, said the company continued to see “divergency in spending between the income groups” last quarter. “The pressure on middle- and low-end-income consumers is still there.”

The widening K

The U.S. economy has been turning more “K”-shaped for decades, with the high-earner cohort doing better and better while others fall further down the economic ladder.

In 1989, the top 10% of U.S. wealth holders already controlled about 61% of the total wealth in the economy, according to Federal Reserve data. Today, it’s about 67%.

While inequality has grown throughout this period, its rate of growth has ebbed and risen.

For example, signs of bifurcation blared louder during the first six months of the Covid-19 pandemic. As the stock market began to recover its early losses, huge parts of the economy remained shut down.

But these signs receded somewhat in the following years, as government stimulus checks and loan-payment forbearance helped boost the fortunes of lower-income households.

Five years on, worrying signals of inequality are roaring back into view. This time, the cracks appear to be hardening.

Average worker pay only barely outpaces inflation — which remains above the Federal Reserve’s 2% target. But the pace of that wage growth is higher for salaried workers than for those paid by the hour, according to data from the Atlanta Federal Reserve.

For nearly all workers, the employment outlook is uncertain, with mass layoffs in the news and AI doing work that people used to do.

“While upper-income consumers are doing great and benefiting from the wealth effect of owning their home and having a stock portfolio, anyone renting or who doesn’t own stocks is not,” said Peter Boockvar, chief investment officer at OnePoint BFG Wealth Partners. “At best, their wages are only keeping up. They’re doing a lot of running in place.”

Two kinds of consumers

The same K shape that characterizes widening income inequality is reflected in consumer spending.

As a result, the volume of companies reporting evidence of a two-tiered economy among their customers surged in recent weeks.

Chipotle CEO Scott Boatwright said the restaurant chain had seen the divide among its customers deepen just in the past few months.

“Earlier this year, as consumer sentiment declined sharply, we saw a broad-based pullback in frequency [of restaurant visits] across all income” levels, he said on Chipotle’s earnings call Wednesday.

“Since then, the gap has widened, with low- to middle-income guests further reducing frequency” of their visits, he said.

Younger adults ages 25 to 35 are being particularly hard hit, said Boatwright. “This group is facing several headwinds, including unemployment, increased student loan repayment and slower real wage growth.”

It’s not just fast-casual dining that’s feeling the loss of lower-income customers.

On Tuesday, the CEO of snack giant Mondelez said the company saw weakness in its cookie and cracker segment last quarter partly due to “value-seeking consumers increasingly turning to discounters and club stores.”

Yet a decline in consumers willing to pay for name-brand snacks like Oreos and Chips Ahoy is only part of the challenge, said chief executive Dirk Van de Put.

“Higher-income consumers increasingly are choosing premium and better-for-you products,” he said, a category that excludes many of Mondelez’s biggest sellers.

The twin issues, he said, were starting to affect the company’s bottom line.

Washington is watching

The growing inequality between a small percentage of consumers and everyone else has caught the attention of the Federal Reserve.

Fed Chair Jerome Powell said he had noticed the surge in consumer-oriented companies reporting split-screen spending patterns.

They “are saying there is a bifurcated economy there, and that consumers at the lower end are struggling and buying less and shifting to lower-cost products,” Powell said Wednesday.

“But at the top, people are spending, at the higher income and wealth,” said Powell. “So, we think there is something there.”

President Donald Trump’s tariffs are part of the equation, too, Fed governor Christopher Waller told CNBC.

Rising prices on a subset of consumer goods hit with tariffs has contributed to a “two-tier” spending scenario where higher-income shoppers had become virtually “price insensitive,” he said.

Meanwhile, lower-income customers will simply “walk out the door” if they see higher prices, Waller said.

The wealthy keep shopping

As lower-income consumers pull back, spending by the top half of the K is increasingly helping to keep the economy humming.

The top 10% of earners account for about half of all consumer spending, according to recent estimates from Moody’s Analytics.

This month, American Express reported that spending by its typically higher-income card holders accelerated by 8%. The number of newly opened “Platinum Card” accounts doubled, partly due to a recent “refresh” of the card’s perks.

Automakers and airlines have likewise expanded their higher-end offerings — with stellar results.

Delta Air Lines now expects sales of premium seats like Comfort-Plus to exceed sales of main cabin seats for the first time, starting next year.

“Premium products used to be loss leaders, and now they’re the highest-margin products,” airline President Glen Hauenstein said on the company’s recent earnings call.

Likewise, Ford and GM both reported booming sales for their largest and most expensive SUVs.

“Today’s auto market is being driven by wealthier households who have access to capital, good loan rates and are propping up the higher end of the market,” Erin Keating, an executive analyst at Cox Automotive, wrote in September.

Beyond consumer spending

Simply looking at consumer behavior may not capture the full financial picture of American households, said Chris Wheat, managing director at JPMorganChase & Co. and president of the the bank’s research institute.

“Just looking at spending, do you really have the full picture of ‘How K-shaped is the K?’” he said. “There are other ways people might be taking advantage of some of the economic growth we’ve been experiencing.”

For example, there are signs that more lower-income households are investing in the stock market than a decade ago.

As of May 2025, the share of people with below-median incomes transferring funds to investment accounts was about five times higher than the average seen between 2010 and 2015, according to data from the JPMorganChase Institute.

Meanwhile, there is evidence that the K may be flattening for the rich.

Upper-income households face new financial pressures today in a rapidly changing labor market that’s witnessing an unprecedented slowdown in the growth of traditionally higher-paying white-collar jobs.

Credit delinquencies among individuals making $150,000 or more have doubled since 2023, according to credit modeler VantageScore.

During the same period, delinquencies for people making between $45,000 and $150,000 rose 58%, and just 17% for borrowers earning less than $45,000.

The key distinction is annual income versus total wealth, said Rikard Bandebo, VantageScore’s chief economist.

Today, the share of jobs being created that are considered high-income has fallen to its lowest level since at least 2015, he said.

This suggests that even a well-paying job is no longer a reliable source of financial stability compared with owning assets like stocks and real estate.

“We are seeing more people in the U.S. economy struggling to make ends meet — and even seeing people in the higher-income range struggling,” said Bandebo.

“It’s not as simple as income being the differentiator here — it’s wealth being more the driver,” he said.

S&P Global Ratings has assigned a so-called junk bond rating to Bitcoin treasury company Strategy (NASDAQ:MSTR).

The ratings firm on Monday assigned Strategy a B- rating with a stable outlook, well below its investment-grade threshold, citing the company’s “high bitcoin concentration, narrow business focus, weak risk-adjusted capitalization, and low U.S. dollar liquidity.”

The rating falls into S&P’s speculative grade. It suggests that the company is “able to service debt for now, but vulnerable to shocks,” VanEck Head of Digital Assets Research Matthew Sigel said in an X post.

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Over the past five years, Strategy has shifted its focus from software to issuing debt to accumulate Bitcoin. This has made its stock a proxy for the digital asset for investors who either cannot access the asset directly or do not want to.

S&P said that while Strategy’s debt obligations were denominated in dollars, the company had low dollar liquidity because it does not generate cash and all excess cash is used for Bitcoin purchases and company operations.

S&P said Strategy’s model depended on the company’s ability to raise capital, including to meet debt obligations. The firm warned that ability could be significantly hampered in a Bitcoin downturn, which could lead to a default or the company selling its Bitcoin holdings at lower prices as a last resort.

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Still, Strategy’s credit rating has been celebrated by some proponents. For example, Strategy Chair Michael Saylor on X said that the company was the first Bitcoin treasury firm to receive a major credit rating.

Meanwhile, BTC Inc. CEO David Bailey said, “The market demand for treasury companies is about to explode.”

S&P said it could upgrade Strategy’s rating if the company improved its dollar liquidity, reduced its reliance on convertible debt and continued to demonstrate a strong ability to raise capital even in Bitcoin downturns.

However, an upgrade is unlikely in the next 12 months, S&P added.

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On September 24, 2025, the Company’s Board of Directors approved the project to convert American Depositary Receipts (ADRs) into ordinary shares listed on the New York Stock Exchange (NYSE).

With this project, TotalEnergies is adapting to the evolution of its shareholder base, which has become increasingly weighted toward North America. In addition, the Conversion will remove ADR frictions, foster access to additional Assets Under Management (AUM) among institutional investment funds and enhance trading liquidity.

The Company has launched on October 30, 2025 the termination of the deposit agreement between TotalEnergies, JP Morgan Chase Bank, N.A. (the depositary), and the ADR holders governing the ADR program.

Upon termination of the deposit agreement, each outstanding ADR will be cancelled and an ordinary share listed on the NYSE will be delivered. The conversion of the ADRs into ordinary shares listed on the NYSE is expected to become effective from December 8, 2025.

General information regarding the listing of TotalEnergies ordinary shares on the NYSE is available online: Expected Timetable and Frequently Asked Questions.