Unveiling the limits of precision in iterative MINFLUX.

In single-molecule microscopy, a big question is how precisely we can estimate the location of a single molecule. Our research shows that by using iterative localisation microscopy and factoring in the prior information, we can boost precision and reduce the number of photons needed. Leveraging the Van Trees inequality aids in determining the optimal precision achievable. Our approach holds promise for wider application in discerning the optimal precision across diverse imaging scenarios, encompassing various illumination strategies, point spread functions and overarching control methodologies.

Targeted axillary dissection reduces residual nodal disease in clinically node- positive breast cancer after neoadjuvant chemotherapy.

BACKGROUND: Any advantage of performing targeted axillary dissection (TAD) compared to sentinel lymph node (SLN) biopsy (SLNB) is under debate in clinically node-positive (cN+) patients diagnosed with breast cancer. Our objective was to assess the feasibility of the removal of the clipped node (RCN) with TAD or without imaging-guided localisation by SLNB to reduce the residual axillary disease in completion axillary lymph node dissection (cALND) in cN+ breast cancer. METHODS: A combined analysis of two prospective cohorts, including 253 patients who underwent SLNB with/without TAD and with/without ALND following NAC, was performed. Finally, 222 patients (cT1-3N1/ycN0M0) with a clipped lymph node that was radiologically visible were analyzed. RESULTS: Overall, the clipped node was successfully identified in 246 patients (97.2%) by imaging. Of 222 patients, the clipped lymph nodes were non-SLNs in 44 patients (19.8%). Of patients in cohort B (n=129) with TAD, the clipped node was successfully removed by preoperative image-guided localisation, or the clipped lymph node was removed as the SLN as detected on preoperative SPECT-CT. Among patients with ypSLN(+) (n=109), no significant difference was found in non-SLN positivity at cALND between patients with TAD and RCN (41.7% vs. 46.9%, p=0.581). In the subgroup with TAD with axillary lymph node dissection (ALND; n=60), however, patients with a lymph node (LN) ratio (LNR) less than 50% and one metastatic LN in the TAD specimen were found to have significantly decreased non-SLN positivity compared to others (27.6% vs. 54.8%, p=0.032, and 22.2% vs. 50%, p=0.046). CONCLUSIONS: TAD by imaging-guided localisation is feasible with excellent identification rates of the clipped node. This approach has also been found to reduce the additional non-SLN positivity rate to encourage omitting ALND in patients with a low metastatic burden undergoing TAD.

Spontaneous tumours in dogs: A clinical and pathomorphological study in Kyrgyzstan.

The aim of this study is to investigate the correlation between the incidence of spontaneous tumours of various origins and the localisation in dogs with sex, breed, and age factors. A total of 360 tumours with various localisation were studied pathomorphologically. Histopathologic data sets from 360 dog tissue samples were processed and statistically examined. A chi-square test of independence was conducted to examine the relationships among the various levels of the specified variables. Logistic regression models were employed for dichotomous outcomes to ascertain the influence of certain explanatory variables on the tumour types. Characteristic pathomorphological changes observed during examination of dogs with oncologic diseases were determined. The most common neoplasms were mammary tumours, accounting for 43% of the cases. The mammary gland tumours were most common in mongrel dogs (25%), with German Shepherds (17.3%), Poodles, Dachshunds, Central Asian Shepherds (6.7% each), and Rottweilers (5.7%) following. The highest frequency of these tumours appeared at 8 years of age, predominantly originating from the ductal epithelium, which represented 46.4% of all the malignant cases.

Intracellular localisation and extracellular release of Y RNA and Y RNA binding proteins.

Cells can communicate via the release and uptake of extracellular vesicles (EVs), which are nano-sized membrane vesicles that can transfer protein and RNA cargo between cells. EVs contain microRNAs and various other types of non-coding RNA, of which Y RNA is among the most abundant types. Studies on how RNAs and their binding proteins are sorted into EVs have mainly focused on comparing intracellular (cytoplasmic) levels of these RNAs to the extracellular levels in EVs. Besides overall transcriptional levels that may regulate sorting of RNAs into EVs, the process may also be driven by local intracellular changes in RNA/RBP concentrations. Changes in extracellular Y RNA have been linked to cancer and cardiovascular diseases. Although the loading of RNA cargo into EVs is generally thought to be influenced by cellular stimuli and regulated by RNA binding proteins (RBP), little is known about Y RNA shuttling into EVs. We previously reported that immune stimulation alters the levels of Y RNA in EVs independently of cytosolic Y RNA levels. This suggests that Y RNA binding proteins, and/or changes in the local Y RNA concentration at EV biogenesis sites, may affect Y RNA incorporation into EVs. Here, we investigated the subcellular distribution of Y RNA and Y RNA binding proteins in activated and non-activated THP1 macrophages. We demonstrate that Y RNA and its main binding protein Ro60 abundantly co-fractionate in organelles involved in EV biogenesis and in EVs. Cellular activation led to an increase in Y RNA concentration at EV biogenesis sites and this correlated with increased EV-associated levels of Y RNA and Ro60. These results suggest that Y RNA incorporation into EVs may be controlled by local intracellular changes in the concentration of Y RNA and their protein binding partners.

Plague and local response in North China, 1900-28.

Breakthroughs in international biomedical science circa 1900 meant that plague could be contained through strict quarantine regulations. These measures were successfully deployed with help from local governments during outbreaks of pneumonic plague in Manchuria (1910-11), Shanxi (1918), and elsewhere in North China. This containment shows the effectiveness of uniting international knowledge and local cooperation in disaster response. Yet, in later outbreaks in similar locations, control measures identical to those instituted a decade earlier were rejected, and plague spread largely unchecked. Historical case studies of the control and spread of infectious disease in North China reveal the complexities of the relationship between global knowledge and its broader, local integration, variation in what constitutes effective 'local' cooperation in adopting international knowledge, and the paramount importance of the locality to the landscape of disaster response. History can reveal critical issues in localisation of disaster response still salient today.

Diasporas as a linchpin in local and international humanitarian action: a case study of the Chinese in Aceh following the 2004 tsunami.

Chinese humanitarian actors have worked frequently with the Chinese diaspora in disaster-affected areas, but little, if any, research has been conducted into the important role of the diaspora in disaster response and humanitarian assistance. This paper investigates what local knowledge the Chinese diaspora has offered to humanitarian actors from the People's Republic of China (PRC), and how this has contributed to their effectiveness. Based on a case study of the semi-autonomous Indonesian province of Aceh in the aftermath of the Indian Ocean tsunami of 2004, this paper argues that the diaspora can serve as a linchpin in local and international humanitarian action. It can do so by strengthening networks and bringing together local ethnic communities, local governments, and the PRC's humanitarian actors, while also offering local knowledge in the form of contextual memory. Such local knowledge may have to be fully utilised to address any underlying ethnic tensions in disaster-affected areas.

Local knowledge as the basis of disaster management and humanitarian assistance.

Recent policy discourse on the localisation of disaster management and humanitarian assistance lacks attention to the culture, history, and traditions of the Global South. This special issue of Disasters argues that it is imperative to recognise the dynamic, interactive, contested, and negotiated nature of local knowledge. Such local knowledge saves lives by enabling responders to situate ad hoc, one-off events such as disasters in the broader and deeper context of community relationships, thereby providing more appropriate and more effective aid. Through the cases of China, Japan, Indonesia, and the Philippines, this special issue examines such dynamic local knowledge using an analytical framework consisting of three manifestations of local knowledge, namely: social capital; contextual historical memories; and adaptation to new ideas. These three manifestations show the ways in which local knowledge creates local capacity, via which local, national, and international disaster respondents can centre their response coordination, and in turn, demonstrate how local capacity reformulates local knowledge.

Phrenitis and the pathology of the mind in western medical thought (fifth century BCE to twentieth century cE).

Phrenitis is ubiquitous in ancient medicine and philosophy. Galen mentions the disease innumerable times, Patristic authors take it as a favourite allegory of human flaws, and no ancient doctor fails to diagnose it and attempt its cure. Yet the nature of this once famous disease has not been properly understood by scholars. My book provides the first full history of phrenitis. In doing so, it surveys ancient ideas about the interactions between body and soul, both in health and in disease. It also addresses ancient ideas about bodily health, mental soundness and moral 'goodness', and their heritage in contemporary psychiatry, offering a chance to reflect critically on contemporary ideas about what it means to be 'insane'.

Enmity then empathy: How militarisation facilitated collaborative but exclusive exchange in Sierra Leone's Ebola response.

In the autumn of 2014, with the 2013-16 West Africa Ebola epidemic spiralling out of control, the United Kingdom announced a bespoke military mission to support-and in some ways lead-numerous Ebola response functions in Sierra Leone. This study examines the nature and effect of the civil-military relationships that subsequently developed between civilian and military Ebola response workers (ERWs). In total, 110 interviews were conducted with key involved actors, and the findings were analysed by drawing on the neo-Durkheimian theory of organisations. This paper finds that stereotypical opposition between humanitarian and military actors helps to explain how and why there was initial cooperative and collaborative challenges. However, all actors were found to have similar hierarchical structures and operations, which explains how and why they were later able to cooperate and collaborate effectively. It also explains how and why civilian ERWs might have served to exclude and further marginalise some local actors.

The emotion of sound target modulates the auditory gaze cueing effect.

The auditory gaze cueing effect (auditory-GCE) is a faster response to auditory targets at an eye-gaze cue location than at a non-cue location. Previous research has found that auditory-GCE can be influenced by the integration of both gaze direction and emotion conveyed through facial expressions. However, it is unclear whether the emotional information of auditory targets can be cross-modally integrated with gaze direction to affect auditory-GCE. Here, we set neutral faces with different gaze directions as cues and three emotional sounds (fearful, happy, and neutral) as targets to investigate how the emotion of sound target modulates the auditory-GCE. Moreover, we conducted a controlled experiment using arrow cues. The results show that the emotional content of sound targets influences the auditory-GCE but only for those induced by facial cues. Specifically, fearful sounds elicit a significantly larger auditory-GCE compared to happy and neutral sounds, indicating that the emotional content of auditory targets plays a modulating role in the auditory-GCE. Furthermore, this modulation appears to occur only at a higher level of social meaning, involving the integration of emotional information from a sound with social gaze direction, rather than at a lower level, which involves the integration of direction and auditory emotion.

Ergothioneine and mitochondria: An important protective mechanism?

Mitochondrial dysfunction is implicated in a wide range of human disorders including many neurodegenerative and cardiovascular diseases, metabolic diseases, cancers, and respiratory disorders. Studies have suggested the potential of l-ergothioneine (ET), a unique dietary thione, to prevent mitochondrial damage and improve disease outcome. Despite this, no studies have definitively demonstrated uptake of ET into mitochondria. Moreover, the expression of the known ET transporter, OCTN1, on the mitochondria remains controversial. In this study, we utilise mass spectrometry to demonstrate direct ET uptake in isolated mitochondria as well as its presence in mitochondria isolated from ET-treated cells and animals. Mitochondria isolated from OCTN1 knockout mice tissues, have impaired but still detectable ET uptake, raising the possibility of alternative transporter(s) which may facilitate ET uptake into the mitochondria. Our data confirm that ET can enter mitochondria, providing a basis for further work on ET in the prevention of mitochondrial dysfunction in human disease.

Proximity Labelling to Quantify Kv7.4 and Dynein Protein Interaction in Freshly Isolated Rat Vascular Smooth Muscle Cells.

Understanding protein-protein interactions is crucial for unravelling subcellular protein distribution, contributing to our understanding of cellular organisation. Moreover, interaction studies can reveal insights into the mechanisms that cover protein trafficking within cells. Although various techniques such as Förster resonance energy transfer (FRET), co-immunoprecipitation, and fluorescence microscopy are commonly employed to detect protein interactions, their limitations have led to more advanced techniques such as the in situ proximity ligation assay (PLA) for spatial co-localisation analysis. The PLA technique, specifically employed in fixed cells and tissues, utilises species-specific secondary PLA probes linked to DNA oligonucleotides. When proteins are within 40 nm of each other, the DNA oligonucleotides on the probes interact, facilitating circular DNA formation through ligation. Rolling-circle amplification then produces DNA circles linked to the PLA probe. Fluorescently labelled oligonucleotides hybridise to the circles, generating detectable signals for precise co-localisation analysis. We employed PLA to examine the co-localisation of dynein with the Kv7.4 channel protein in isolated vascular smooth muscle cells from rat mesenteric arteries. This method enabled us to investigate whether Kv7.4 channels interact with dynein, thereby providing evidence of their retrograde transport by the microtubule network. Our findings illustrate that PLA is a valuable tool for studying potential novel protein interactions with dynein, and the quantifiable approach offers insights into whether these interactions are changed in disease.

Augmented-reality-based surgical navigation for endoscope retrograde cholangiopancreatography: A phantom study.

BACKGROUND: Endoscope retrograde cholangiopancreatography is a standard surgical treatment for gallbladder and pancreatic diseases. However, surgeons is at high risk and require sufficient surgical experience and skills. METHODS: (1) The simultaneous localisation and mapping technique to reconstruct the surgical environment. (2) The preoperative 3D model is transformed into the intraoperative video environment to implement the multi-modal fusion. (3) A framework for virtual-to-real projection based on hand-eye alignment. For the purpose of projecting the 3D model onto the imaging plane of the camera, it uses position data from electromagnetic sensors. RESULTS: Our AR-assisted navigation system can accurately guide physicians, which means a distance of registration error to be restricted to under 5 mm and a projection error of 5.76 ± 2.13, and the intubation procedure is done at 30 frames per second. CONCLUSIONS: Coupled with clinical validation and user studies, both the quantitative and qualitative results indicate that our navigation system has the potential to be highly useful in clinical practice.

Tumour-specific phosphorylation of serine 419 drives alpha-enolase (ENO1) nuclear export in triple negative breast cancer progression.

BACKGROUND: The glycolytic enzyme alpha-enolase is a known biomarker of many cancers and involved in tumorigenic functions unrelated to its key role in glycolysis. Here, we show that expression of alpha-enolase correlates with subcellular localisation and tumorigenic status in the MCF10 triple negative breast cancer isogenic tumour progression model, where non-tumour cells show diffuse nucleocytoplasmic localisation of alpha-enolase, whereas tumorigenic cells show a predominantly cytoplasmic localisation. Alpha-enolase nucleocytoplasmic localisation may be regulated by tumour cell-specific phosphorylation at S419, previously reported in pancreatic cancer. RESULTS: Here we show ENO1 phosphorylation can also be observed in triple negative breast cancer patient samples and MCF10 tumour progression cell models. Furthermore, prevention of alpha-enolase-S419 phosphorylation by point mutation or a casein kinase-1 specific inhibitor D4476, induced tumour-specific nuclear accumulation of alpha-enolase, implicating S419 phosphorylation and casein kinase-1 in regulating subcellular localisation in tumour cell-specific fashion. Strikingly, alpha-enolase nuclear accumulation was induced in tumour cells by treatment with the specific exportin-1-mediated nuclear export inhibitor Leptomycin B. This suggests that S419 phosphorylation in tumour cells regulates alpha-enolase subcellular localisation by inducing its exportin-1-mediated nuclear export. Finally, as a first step to analyse the functional consequences of increased cytoplasmic alpha-enolase in tumour cells, we determined the alpha-enolase interactome in the absence/presence of D4476 treatment, with results suggesting clear differences with respect to interaction with cytoskeleton regulating proteins. CONCLUSIONS: The results suggest for the first time that tumour-specific S419 phosphorylation may contribute integrally to alpha-enolase cytoplasmic localisation, to facilitate alpha-enolase's role in modulating cytoskeletal organisation in triple negative breast cancer. This new information may be used for development of triple negative breast cancer specific therapeutics that target alpha-enolase.

Mendelian randomization combined with multi-omics explores the relationship between heart failure and cancer.

Background: Whether there is an association between HF (HF) and cancer has not been conclusively established, and it is not clear whether patients with cancer can share similar hospitalization strategies and outcomes with patients with HF. Methods: Genome-wide association summary statistics were performed using a two-sample Mendelian randomization (MR) method for HF patients and cancer patients from the GWAS directory, with co-localization and Summary Data-Based Mendelian Randomization (SMR) analyses to identify HF-associated genes, and transcriptomic analyses to analyze the roles of these genes in the clinical diagnosis and targeted therapies of multiple cancer types. Results: Two-sample MR analysis showed that increased risk of HF was associated with decreased risk of cervical, brain, breast, colorectal, lung, and skin cancers, and co-localization combined with SMR analysis identified ABO and SURF1 as HF-associated genes, and transcriptomic analyses showed that ABO is a risk factor for HF and a protective factor against cancer, whereas SURF1 is a protective factor against HF and a protective factor against cancer. Conclusion: There was no causal relationship between heart failure and cancers (Cervical, brain, breast, colorectal, lung and skin cancers) risk factors, however there was a trend toward a negative causal relationship between heart failure and cancers (Cervical, brain, breast, colorectal, lung and skin cancers) occurrence.

Nonpalpable implant removals at centre of experience in France: a cohort study.

OBJECTIVE(S): Management and localisation strategies to remove nonpalpable contraceptive implants may be difficult. We aimed to evaluate our imaging modalities to identify deep implant and patient outcomes related to removal. STUDY DESIGN: In this retrospective study, we reviewed all cases referred to our specialised centre for nonpalpable contraceptive implants from January 2018 to August 2022. RESULTS: Out of the cohort studied, 47 female subjects exhibited nonpalpable implants. The implant was nonpalpable for thirty-six patients (76,6%) immediately after the insertion whereas it was not palpable several months after the insertion for eleven patients (23.4%). Twelve patients (25.5%) had one or more failed removal attempts before referral.All 47 implants were successfully visualised via ultrasound in the upper arm: 40 implants (85.1%) were located in the subdermal tissue, 4 (8.5%) were intrafascial and 3 (6.4%) were intramuscular. Depth of the implant was 4.0 mm [1.7 - 12.0]. No clinical factors were statistically associated with differences in depth or location (subdermal vs subfascial). Removal procedures were mainly under local anaesthesia in 74.5% of cases in an outpatient setting. There were two Clavien-Dindo grade 1 complications (one case of cutaneous scar dehiscence and one transient postoperative neuropathic complaint in the upper arm resolved within 3 months under analgetics). CONCLUSIONS: Identification of deep implants requires following the ultrasound modality protocol. Ultrasound detection makes easy and safe implant removal. Training programs for the insertion as well as for the removal of correct and incorrect inserted implants should be continued and developed all around the world.

A Na+ channel receptor of FMRFamide in the cephalopod Sepiella japonica: Identification, characterisation, and expression profiling during different stages of gonadal development.

FMRFamide, a member of the neuropeptide family, is involved in numerous physiological processes. FMRFamide-activated sodium channels (FaNaCs) are a family of non-voltage-gated, amiloride-sensitive, Na+-selective channels triggered by the neuropeptide FMRFamide. In the present study, the full-length cDNA of the FaNaC receptor of Sepiella japonica (SjFaNaC) was cloned. The cDNA of SjFaNaC was 3004 bp long with an open reading frame (ORF) of 1812 bp, encoding 603 amino acid residues with no signal peptide at the N-terminus. Sequence analysis indicated that SjFaNaC shared a high identity with other cephalopods FaNaCs and formed a sister clade with bivalves. The protein structure was predicted using SWISS-MODEL with AcFaNaC as the template. Quantitative real-time PCR (qRT-PCR) revealed that SjFaNaC transcripts were highly expressed in both female and male reproductive organs, as well as in the optic lobe and brain of the central nervous system (CNS). Results of in situ hybridisation (ISH) showed that SjFaNaC mRNA was mainly distributed in the medulla and deep retina of the optic lobe and in both the supraesophageal and subesophageal masses of the brain. Subcellular localisation indicated that the SjFaNaC protein was localised intracellularly and on the cell surface of HEK293T cells. In summary, these findings may lay the foundation for future exploration of the functions of SjFaNaC in cephalopods.

Ultrafast 3-D Transcutaneous Super Resolution Ultrasound Using Row-Column Array Specific Coherence-Based Beamforming and Rolling Acoustic Sub-aperture Processing: In Vitro, in Rabbit and in Human Study.

OBJECTIVE: This study aimed to realise 3-D super-resolution ultrasound imaging transcutaneously with a row-column array which has far fewer independent electronic channels and a wider field of view than typical fully addressed 2-D matrix arrays. The in vivo image quality of the row-column array is generally poor, particularly when imaging non-invasively. This study aimed to develop a suite of image formation and post-processing methods to improve image quality and demonstrate the feasibility of ultrasound localisation microscopy using a row-column array, transcutaneously on a rabbit model and in a human. METHODS: To achieve this, a processing pipeline was developed which included a new type of rolling window image reconstruction, which integrated a row-column array specific coherence-based beamforming technique with acoustic sub-aperture processing. This and other processing steps reduced the 'secondary' lobe artefacts, and noise and increased the effective frame rate, thereby enabling ultrasound localisation images to be produced. RESULTS: Using an in vitro cross tube, it was found that the procedure reduced the percentage of 'false' locations from ∼26% to ∼15% compared to orthogonal plane wave compounding. Additionally, it was found that the noise could be reduced by ∼7 dB and the effective frame rate was increased to over 4000 fps. In vivo, ultrasound localisation microscopy was used to produce images non-invasively of a rabbit kidney and a human thyroid. CONCLUSION: It has been demonstrated that the proposed methods using a row-column array can produce large field of view super-resolution microvascular images in vivo and in a human non-invasively.

Prevalence and characterisation of band-shaped tail lesions in Holstein cows.

The aim of the study was to characterise and determine the prevalence of band-shaped tail lesions in Holstein cows. Lesions were present either as wounds or by epithelised granulation/connective tissue formations. Both types were characterised by a median localisation 7 cm from the tip of the tail, and they occurred on the dorsal aspect of the tail. From here they encircled the tail either completely or in varying degrees, and they were often present as isolated lesions (93%). The prevalence of band-shaped tail lesions was found to be 25% among 2099 cows examined in 16 Danish Holstein herds with a variation from 18 to 40% between herds. In the herds, the wound lesions and the connective tissue formations accounted for 22% and 78% of all band-shaped tail lesions, respectively. Among 458 Holstein cows examined at an abattoir the prevalence of band-shaped tail lesions was 23%, i.e. similar to the prevalence within the herds. At the abattoir the share of band-shaped wound lesions was 67% and the band-shaped connective tissue formation 33%. Associations between the occurrence of band-shaped tail lesions and parity and lack of the tail tip were observed.

Membrane Association of the Short Transglutaminase Type 2 Splice Variant (TG2-S) Modulates Cisplatin Resistance in a Human Hepatocellular Carcinoma (HepG2) Cell Line.

Hepatocellular carcinoma (HCC) is a heterogeneous malignancy with complex carcinogenesis. Although there has been significant progress in the treatment of HCC over the past decades, drug resistance to chemotherapy remains a major obstacle in its successful management. In this study, we were able to reduce chemoresistance in cisplatin-resistant HepG2 cells by either silencing the expression of transglutaminase type 2 (TG2) using siRNA or by the pre-treatment of cells with the TG2 enzyme inhibitor cystamine. Further analysis revealed that, whereas the full-length TG2 isoform (TG2-L) was almost completely cytoplasmic in its distribution, the majority of the short TG2 isoform (TG2-S) was membrane-associated in both parental and chemoresistant HepG2 cells. Following the induction of cisplatin toxicity in non-chemoresistant parental cells, TG2-S, together with cisplatin, quickly relocated to the cytosolic fraction. Conversely, no cytosolic relocalisation of TG2-S or nuclear accumulation cisplatin was observed, following the identical treatment of chemoresistant cells, where TG2-S remained predominantly membrane-associated. This suggests that the deficient subcellular relocalisation of TG2-S from membranous structures into the cytoplasm may limit the apoptic response to cisplatin toxicity in chemoresistant cells. Structural analysis of TG2 revealed the presence of binding motifs for interaction of TG2-S with the membrane scaffold protein LC3/LC3 homologue that could contribute to a novel mechanism of chemotherapeutic resistance in HepG2 cells.