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This study describes the congenital goiter in an alpaca (Vicugna pacos) fetus aborted in November 2021 with the clinical and pathological findings in the dam that was found dead on the farm three weeks after a miscarriage. The dam was a black coat alpaca bred in the Netherlands, imported in Italy in January 2021, and housed in a farm of central Italy for breeding purposes. Signalment and clinical data on dam and fetus were collected from the farmer and referring veterinarian. Necropsies and histopathological investigations on mother and conceptus were conducted at the Department of Veterinary Medicine, University of Perugia. The fetus was aborted at the 5th gestational month and at necropsy revealed bilateral thyroid gland enlargement. Microscopically the thyroid gland presented follicular cell hypertrophy and hyperplasia, follicles greatly varying in size, with macrofollicles lined by attenuated to atrophic epithelium and distended by eosinophilic colloid (colloid goiter). The dam presented a history of anorexia, wasting, dullness, severe weakness, incoordination, recurrent seizures, and, most recently, abortion. At necropsy, a cerebral focal loss of substance was found at the right parietal cortex, which histologically corresponded to laminar ischemic neuronal necrosis; most importantly, histological examination of cerebellum showed multifocal necrosis of the Purkinje cells associated with severe depletion of the granular layer neurons. These cerebellar findings in the dam combined with goiter in the conceptus suggest organomercurial poisoning as the likely cause of abortion followed by mother's death. Mercury poisoning is hypothesized as the possible link between mother's and fetus' lesions., Competing Interests: Declarations. Animal ethics approval: . Not applicable. Clinical trial number: Not applicable. Competing interests: The authors declare no competing interests., (© 2025. The Author(s), under exclusive licence to Springer Nature B.V.)

Knowing mechanisms that facilitate the emergence of post-glacial ecosystems is urgently required given rapid recent glacial retreat in high latitude and high elevation regions. We examined the effect of nutrient hotspots created via communal dung deposition by wild, native Andean camelids on soil abiotic and biotic properties and plant cover in the rapidly deglaciating Cordillera Vilcanota, southeastern Peru. Animal-modified proglacial soils were significantly enriched in all measured edaphic properties compared to reference glacial-till soils of the same age adjacent to animal-modified soil patches. Organic matter composition, soil moisture, available inorganic nitrogen, and plant cover were nearly zero in glacial-moraine reference soils, but were at least one order of magnitude greater in animal-modified soils. Likewise, DNA concentrations were almost two orders of magnitude higher in modified soils (23 ± 9 µg DNA g soil -1 ) compared to reference soils (0.6 ± 0.3 µg DNA g soil -1 ). Animal-modified soil microbial community composition differed significantly from reference soils for both prokaryotes and eukaryotes, and eukaryote ASV richness was significantly higher in camelid latrines than in controls. Nutrient transfer into glacier forefields by native camelids shortcuts a 100+ year lag between glacier retreat and primary succession. Our results suggest that nutrient transfer into glacier forefields by wild, native animals may be an important, natural mechanism by which tropical Andean species can expand upslope at a pace relevant to climate change., Competing Interests: Declarations. Competing interests: The authors declare no competing interests., (© 2024. The Author(s).)

In the high altitudes of the Andes, llama breeders shear the fiber from their animals, obtaining fleeces for many purposes. Dehairing the fleece of these animals is a viable alternative to improving the quality and value of the fleece. The study examined the attributes of fiber quality and pilose follicle of dehaired and non-dehaired fleece from Ch'aku llamas and the relationship among these characteristics. From 80 animals, 480 fleece samples (240 dehaired and 240 non-dehaired), and 198 skin biopsies of dehaired specimens were collected. The data was analyzed using a model that contain fleece type effect, age group effect, sex effect, body region effect, and their respective interactions. Also, the Spearman correlation between fiber quality attributes and pilose follicle traits was calculated. The findings showed that non-dehaired fiber had a diameter of 22.01 ± 0.17 µm, while dehaired fiber measured 20.85 ± 0.16 µm. Removing the hair from the fiber resulted in a decrease in the average fiber diameter, standard deviation, and coefficient of variation while increasing the comfort factor and curvature index. The age group had an impact on primary follicles, secondary follicles, follicular density, and follicular index. Furthermore, gender influenced secondary follicles and follicular density, whereas body region affected secondary follicles and follicular density. Negative correlations were observed between mean of follicular density (MFoD) and mean fiber diameter (MFD), as well as curvature ratio (CRV), whereas a positive correlation was found between mean of follicular index (MFoI) and MFoD, and a negative correlation was noted between MFoI and CRV. It is concluded that the dehairing of the Ch'aku llama fiber improves the quality of the fleece., Competing Interests: Declarations. Ethical statement: All animals were handled and treated according to the guidelines approved by the Ethics Committee for Research (R-093–2018-CU-UNAMBA) at Universidad Nacional del Micaela Bastidas de Apurímac (Abancay, Apurímac, Perú). Conflict of interest: The authors declare that they have no conflicts of interest., (© 2024. The Author(s), under exclusive licence to Springer Nature B.V.)

In pathological examinations, gastric ulcers are often detected in South American camelids. The aetiology cannot be clarified in every case. However, tumour-related gastric ulcers are observed repeatedly. This study reports clinical, laboratory and pathological findings in six alpacas, three each with adenocarcinoma- and squamous cell carcinoma-associated gastric ulcers. Clinically they were presented with general symptoms like emaciation, anorexia and recumbency. Laboratory findings of these animals were non-specific. All animals were at least nine years old. The adenocarcinomas as well as the squamous cell carcinomas were metastasised in various organs, especially in the lymph nodes. Two adenocarcinoma-associated gastric ulcers were perforated. In summary, squamous cell carcinomas and adenocarcinomas can lead to ulcerative lesions in the gastrointestinal tract. Although neoplasms are rare overall, they should be considered as a possible differential diagnosis in the case of non-specific symptoms. In particular, older animals seem to be affected more frequently., Competing Interests: Competing interests: The authors declare no competing interests., (© 2024. The Author(s).)

In head and neck squamous cell carcinoma (HNSCC), the mesenchymal epithelial transition (MET) receptor drives cancer growth, proliferation, and metastasis. MET is known to be overexpressed in HNSCC and, therefore, is an appealing therapeutic target. In this study, we evaluated MET expression in patients with HNSCC and investigated the potential imaging application of a novel MET-binding single-domain camelid antibody using positron emission tomography/computed tomography (PET/CT) in a preclinical MET-expressing HNSCC model. Multiplex immunostaining for MET protein performed on a tissue microarray from 203 patients with HNSCC found 86% of patients to have MET expression, with 14% having high expression and 53% having low MET expression. Using The Cancer Genome Atlas (TCGA) database, high MET RNA expression was associated with worse progression-free survival and overall survival in patients with HPV-negative HSNCC. Utilizing flow cytometry and immunofluorescence, our novel camelid antibody fused to a human IgG Fc chain (1E7-Fc) showed high binding affinity and specificity to high MET-expressing Detroit 562 cells but not to low MET-expressing HNSCC cells. The efficacy and biodistribution of [ 89 Zr]Zr-1E7-Fc as a PET imaging agent was then investigated in a MET-expressing head and neck xenograft model. [ 89 Zr]Zr-1E7-Fc rapidly localized and showed high tumor uptake in Detroit 562 xenografts (8.4% ID/g at 72 h postinjection), with rapid clearance from the circulatory system (2.7 tumor-to-blood radioactivity ratio at 72 h postinjection). Our preclinical data suggest that the camelid antibody 1E7-Fc could be a potential theranostic agent for HNSCC. Further investigations are warranted to confirm these findings in patients and to evaluate 1E7-Fc as an imaging agent and platform to deliver radionuclide or drug therapy to MET-driven cancers.

A 1-year-old alpaca mare was presented due to lateral deviation of the cervical spine which had been present for a period of 2 weeks. Based on the previous report, a traumatic event was assumed. Feed and water intake were unremarkable, the gait was altered due to slight ataxic movements in all 4 limbs. The animal showed no further neurological deficits on initial presentation. Radiographs revealed a displacement of the cervical vertebrae to the left as part of a subluxation of the articular processes of the cervical vertebrae C4 and C5. During the course of conservative treatment with pain medication, neurologic signs including ataxia and I: mpaired balance developed in spite of unremarkable food intake. Subsequently, the animal died at a time when it was not under observation.Pathologic examination revealed degenerative and inflammatory alterations as well as hemorrhages in the cervical medulla at the level of the axial deviation of the cervical spine. Under consideration of the patient's history, the findings may most likely be seen as the result of blunt trauma and represent the cause of the neurologic signs.This is the first case description of a subluxation of the cervical vertebral column in an alpaca in Germany that has been evaluated in detail., Competing Interests: Die Autoren bestätigen, dass kein Interessenkonflikt besteht., (Thieme. All rights reserved.)

A 14-y-old intact female llama ( Lama glama ) was presented for evaluation of a right maxillary swelling of 3-mo duration. Clinically, the animal had mild nasal discharge, abnormal retropulsion of the right eye, and moderate gingival disease. An incisional biopsy of the maxillary mass revealed pleomorphic and mitotically active neoplastic spindle-to-stellate cells organized in haphazard lacunae embedded in abundant chondroid matrix. Given the poor prognosis, euthanasia was elected. Postmortem examination and sectioning of the head exposed a large solid, white, firm mass that vastly expanded the right infraorbital region, extending to the maxilla, effacing the right nasal conchae and ipsilateral zygomatic bone. Collectively, postmortem dissection, cytology, and histopathology of the primary mass supported a diagnosis of sinonasal chondrosarcoma. To our knowledge, this entity had not been reported previously in this species and should be considered a differential for facial deformities in New World camelids., Competing Interests: Declaration of conflicting interestsThe authors declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.

A 5-year-old female alpaca was presented with respiratory distress and lethargy. Thoracic radiographs revealed a cranioventrally distributed alveolar pattern, caudodorsal bronchial pattern, cardiomegaly, increased soft tissue opaque content in the ventral thorax, and rounded soft tissue opaque structures craniodorsal to the carina. Cardiac gated CT demonstrated a patent ductus arteriosus, ventricular septal defect, complete left atrioventricular valve atresia, partial anomalous venous connections from the cranial pulmonary veins to the azygous and cranial vena cava, severe right-sided cardiomegaly, pleural and peritoneal fluid, and severe hepatic congestion. These findings were confirmed with necropsy., (© 2024 American College of Veterinary Radiology.)

SARS-CoV-2 and its variants continue to pose a significant threat to public health. Nanobodies (Nbs) that inhibit the interaction between the receptor-binding domain (RBD) of the spike protein and the host cell receptor angiotensin-converting enzyme 2 (ACE2) are promising drug candidates. In this study, we report the discovery and structural characterization of a potent Nb that targets the RBD. By screening a phage display alpaca naive Nbs library using the RBD as bait, we identified sixteen candidate Nbs. Of these, nine exhibited nanomolar to micromolar binding affinity and strong neutralizing activity against pseudotyped SARS-CoV-2 viruses, with NbS4 showing the highest neutralization potency. The crystal structure of the SARS-CoV-2 RBD in complex with NbS4 revealed that this Nb binds to a site partially overlapping the ACE2 binding region. Importantly, the key binding residues of NbS4 in the RBD are conserved across most known variants, making it a promising candidate for COVID-19 treatment., Competing Interests: Declaration of competing interest The authors have no conflicts of interest to declare., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Antimicrobial resistance is emerging as the new healthcare crisis necessitating the development of newer classes of drugs using non-traditional approaches. Pseudomonas aeruginosa, one of the most common pathogens involved in nosocomial infections, is extremely difficult to treat even with the last resort frontline drug, the carbapenems. As the pathogen has the ability to acquire resistance to new small-molecule antibiotics, being deployed, a novel biological approach has been tried using antibody fragments in combination with carbapenems and β-lactams as adjunct therapy for an enduring solution to the problem. In this study, we developed a camelid antibody fragment (VHH) library against P. aeruginosa and isolated a highly potent hit, PsC23. Mass spectrometry identified the target to be a component of the C4-dicarboxylate transporter that feeds metabolites to the glyoxylate shunt particularly under conditions of oxidative stress. PsC23 is bacteriostatic at a concentration of 1.66 µM (25 µg ml -1 ) and shows a synergistic effect with both the classes of drugs at an effective concentration of 100-200 nM (1.5-3.0 µg ml -1 ) when co administered with them. In combination with meropenem the VHH completely cleared the infection from a neutropenic mouse with a carbapenem-resistant P. aeruginosa systemic infection. Blocking the glyoxylate shunt by PsC23 resulted in disruption of energy transduction due to a respiratory shift to the oxygen-depleted TCA cycle causing inhibition of efflux and increased free radical generation from carbapenems and β-lactams exerting a strong bactericidal effect that reversed the resistance to multiple unrelated drugs., (© 2024. The Author(s), under exclusive licence to the Japan Antibiotics Research Association.)

South American camelids (SAC) are gaining popularity for various purposes, including fiber production, trekking, and companionship. High abortion rates pose a significant health issue in SAC herds, leading to substantial economic losses for breeders. Often, the causes of these abortions remain unidentified. This review provides a comprehensive summary of the known infectious and non-infectious causes of abortions in SAC., Competing Interests: Declaration of Competing Interest The authors declare no potential conflicts of interest., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

Naja atra , the Chinese cobra, is a major cause of snake envenomation in Asia, causing hundreds of thousands of clinical incidents annually. The current treatment, horse serum-derived antivenom, has unpredictable side effects and presents manufacturing challenges. This study focused on developing new-generation snake venom antidotes by using microbial phage display technology to derive nanobodies from an alpaca immunized with attenuated N. atra venom. Following confirmation of the immune response in the alpaca, we amplified V H H genes from isolated peripheral blood mononuclear cells and constructed a phage display V H H library of 1.0 × 10 7 transformants. After four rounds of biopanning, the enriched phages exhibited increased binding activity to N. atra venom. Four nanobody clones with high binding affinities were selected: aNAH1, aNAH6, aNAH7, and aNAH9. Specificity testing against venom from various snake species, including two Southeast Asian cobra species, revealed nanobodies specific to the genus Naja . An in vivo mouse venom neutralization assay demonstrated that all nanobodies prolonged mouse survival and aNAH6 protected 66.6% of the mice from the lethal dosage. These findings highlight the potential of phage display-derived nanobodies as valuable antidotes for N. atra venom, laying the groundwork for future applications in snakebite treatment.IMPORTANCEChinese cobra venom bites present a formidable medical challenge, and current serum treatments face unresolved issues. Our research applied microbial phage display technology to obtain a new, effective, and cost-efficient treatment approach. Despite interest among scientists in utilizing this technology to screen alpaca antibodies against toxins, the available literature is limited. This study makes a significant contribution by introducing neutralizing antibodies that are specifically tailored to Chinese cobra venom. We provide a comprehensive and unbiased account of the antibody construction process, accompanied by thorough testing of various nanobodies and an assessment of cross-reactivity with diverse snake venoms. These nanobodies represent a promising avenue for targeted antivenom development that bridges microbiology and biotechnology to address critical health needs., Competing Interests: The authors declare no conflict of interest.

The excessive deposition of fibrillar collagens is a hallmark of fibrosis. Collagen fibril formation requires proteolytic maturations by Procollagen N- and C-proteinases (PNPs and PCPs) to remove the N- and C-propeptides which maintain procollagens in the soluble form. Procollagen C-Proteinase Enhancer-1 (PCPE-1, a glycoprotein composed of two CUB domains and one NTR domain) is a regulatory protein that activates the C-terminal processing of procollagens by the main PCPs. It is often up-regulated in fibrotic diseases and represents a promising target for the development of novel anti-fibrotic strategies. Here, our objective was to develop the first antagonists of PCPE-1, based on the nanobody scaffold. Using both an in vivo selection through the immunization of a llama and an in vitro selection with a synthetic library, we generated 18 nanobodies directed against the CUB domains of PCPE1, which carry its enhancing activity. Among them, I5 from the immune library and H4 from the synthetic library have a high affinity for PCPE-1 and inhibit its interaction with procollagens. The crystal structure of the complex formed by PCPE-1, H4 and I5 showed that they have distinct epitopes and enabled the design of a biparatopic fusion, the diabody diab-D1. Diab-D1 has a sub-nanomolar affinity for PCPE-1 and is a potent antagonist of its activity, preventing the stimulation of procollagen cleavage in vitro. Moreover, Diab-D1 is also effective in reducing the proteolytic maturation of procollagen I in cultures of human dermal fibroblasts and hence holds great promise as a tool to modulate collagen deposition in fibrotic conditions., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: ‘DV is the former CEO of NVH Medicinal. PL, SVLG, CM and DV have filed a patent regarding the nanobodies described in this study (PCT/FR2023/050369). The authors declare that they have no other competing financial interests’., (Copyright © 2024 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

Fluorescence molecular imaging plays a vital role in image-guided surgery. In this context, the urokinase plasminogen activator receptor (uPAR) is an interesting biomarker enabling the detection and delineation of various tumor types due to its elevated expression on both tumor cells and the tumor microenvironment. In this study, anti-uPAR Nanobodies (Nbs) are generated through llama immunization with human and murine uPAR protein. Extensive in vitro characterization and in vivo testing with radiolabeled variants are conducted to assess their pharmacokinetics and select lead compounds. Subsequently, the selected Nbs are converted into fluorescent agents, and their application for fluorescence-guided surgery is evaluated in various subcutaneous and orthotopic tumor models. The study yields a panel of high-affinity anti-uPAR Nbs, showing specific binding across multiple types of cancer cells in vitro and in vivo. Lead fluorescently-labeled compounds exhibit high tumor uptake with high contrast at 1 h after intravenous injection across all assessed uPAR-expressing tumor models, outperforming a non-targeting control Nb. Additionally, rapid and accurate tumor localization and demarcation are demonstrated in an orthotopic human glioma model. Utilizing these Nbs can potentially enhance the precision of surgical tumor resection and, consequently, improve survival rates in the clinic., (© 2024 The Authors. Advanced Science published by Wiley‐VCH GmbH.)

Opioid use disorders (OUD) and overdoses are ever-evolving public health threats that continue to grow in incidence and prevalence in the United States and abroad. Current treatments consist of opioid receptor agonists and antagonists, which are safe and effective but still suffer from some limitations. Murine and humanized monoclonal antibodies (mAb) have emerged as an alternative and complementary strategy to reverse and prevent opioid-induced respiratory depression. To explore antibody applications beyond traditional heavy-light chain mAbs, we identified and biophysically characterized a novel single-domain antibody specific for fentanyl from a camelid variable-heavy-heavy (VHH) domain phage display library. Structural data suggested that VHH binding to fentanyl was facilitated by a unique domain-swapped dimerization mechanism, which accompanied a rearrangement of complementarity-determining region loops leading to the formation of a fentanyl-binding pocket. Structure-guided mutagenesis further identified an amino acid substitution that improved the affinity and relaxed the requirement for dimerization of the VHH in fentanyl binding. Our studies demonstrate VHH engagement of an opioid and inform on how to further engineer a VHH for enhanced stability and efficacy, laying the groundwork for exploring the in vivo applications of VHH-based biologics against OUD and overdose., Competing Interests: Conflict of interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: A. M. L, J. P. G, M. P., C. B., and D. H. are co-inventors on a provisional patent application USPTO--231220--09824.412., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

To expand the knowledge about common diseases in llamas and alpacas in Germany, a screening of the cases of South American camelids presented at the Clinic for Swine and Small Ruminants of the University of Veterinary Medicine Hannover, Germany from 2005 to the end of November 2021 was performed. A retrospective evaluation of necropsy reports from this period was conducted. Overall, necropsy reports were evaluated from 187 alpacas, 35 llamas and one vicuña (n = 223). A total of 50.2% of the dissected animals were thin or cachectic. Pathological alterations of the gastrointestinal tract were the most common findings (44.8%). In addition, liver changes were recorded, most frequently in adult animals. In contrast, diseases of the respiratory tract and the nervous system were found more frequently in juvenile animals. This study provides an overview of common pathologies in South American camelids in Germany and thus may help to recognise different disease symptoms at an early stage., (© 2024. The Author(s).)

The classical amyloid cascade hypothesis postulates that the aggregation of amyloid plaques and the accumulation of intracellular hyperphosphorylated Tau tangles, together, lead to profound neuronal death. However, emerging research has demonstrated that soluble amyloid-β oligomers (SAβOs) accumulate early, prior to amyloid plaque formation. SAβOs induce memory impairment and disrupt cognitive function independent of amyloid-β plaques, and even in the absence of plaque formation. This work describes the development and characterization of a novel anti-SAβO (E3) nanobody generated from an alpaca immunized with SAβO. In-vitro assays and in-vivo studies using 5XFAD mice indicate that the fluorescein (FAM)-labeled E3 nanobody recognizes both SAβOs and amyloid-β plaques. The E3 nanobody traverses across the blood-brain barrier and binds to amyloid species in the brain of 5XFAD mice. Imaging of mouse brains reveals that SAβO and amyloid-β plaques are not only different in size, shape, and morphology, but also have a distinct spatial distribution in the brain. SAβOs are associated with neurons, while amyloid plaques reside in the extracellular matrix. The results of this study demonstrate that the SAβO nanobody can serve as a diagnostic agent with potential theragnostic applications in Alzheimer's disease., (© 2024. The Author(s).)

Proinflammatory T-lymphocytes recruited into the brain and spinal cord mediate multiple sclerosis (MS) and currently there is no cure for MS. IFN-γ-producing Th1 cells induce ascending paralysis in the spinal cord while IL-17-producing Th17 cells mediate cerebellar ataxia. STAT1 and STAT3 are required for Th1 and Th17 development, respectively, and the simultaneous targeting of STAT1 and STAT3 pathways is therefore a potential therapeutic strategy for suppressing disease in the spinal cord and brain. However, the pharmacological targeting of STAT1 and STAT3 presents significant challenges because of their intracellular localization. We have developed a STAT-specific single-domain nanobody (SBT-100) derived from camelids that targets conserved residues in Src homolog 2 (SH2) domains of STAT1 and STAT3. This study investigated whether SBT-100 could suppress experimental autoimmune encephalomyelitis (EAE), a mouse model of MS. We show that SBT-100 ameliorates encephalomyelitis through suppressing the expansion of Th17 and Th1 cells in the brain and spinal cord. Adoptive transfer experiments revealed that lymphocytes from SBT-100-treated EAE mice have reduced capacity to induce EAE, indicating that the immunosuppressive effects derived from the direct suppression of encephalitogenic T-cells. The small size of SBT-100 makes this STAT-specific nanobody a promising immunotherapy for CNS autoimmune diseases, including multiple sclerosis.

Poisoning is often suspected to be the origin of disease in South American camelids (SACs) by owners, but only in a few cases this assumption can be confirmed. In small ruminants, rhododendron poisoning is a common emergency for livestock veterinarians. However, this condition has rarely been reported in SACs so far. This paper provides information regarding clinical findings, hematology, clinical chemistry, and treatment of four alpacas after presumed intake of rhododendron leaves including pathological findings of one of the animals. Rhododendron leaves contain grayanatoxins that lead to hyperpolarization of excitable cells. Clinical signs that were observed in the presented alpacas comprised: salivation, dehydration, decreased motility of compartment 1, uncoordinated regurgitation, and cardiac arrhythmia. Clinical chemistry revealed that rhododendron poisoning was associated with metabolic acidosis and azotaemia, hyponatremia and hyperkalemia. Most striking macroscopic and histopathological findings included gastric ulceration, and renal infarcts along with inflammatory changes. Leaves of Rhododendron spp. were identified in the forestomach content of this animal. Affected animals were treated symptomatically as there is no specific antidote in rhododendron poisoning. This included parenteral rehydration, treatment of metabolic acidosis (infusion of sodium bicarbonate solution), and oral administration of activated charcoal to bind potential toxins. In addition, antibiotic treatment might be necessary to prevent aspiration pneumonia in case of uncoordinated regurgitation. Of the four animals, the worst affected alpaca was euthanized, one had minimal signs and two responded to supportive care and recovered. In conclusion, rhododendron poisoning might be fatal for alpacas in individual cases and therefore rhododendron bushes should not be placed in the habitat of SACs., (© 2024. The Author(s).)

Single-domain V H H antibody is regarded as one of the promising antibody classes for therapeutic and diagnostic applications. V H H antibodies have amino acids in framework region 2 that are distinct from those in conventional antibodies, such as the Val37Phe/Tyr (V37F/Y) substitution. Correlations between the residue type at position 37 and the conformation of the CDR3 in V H H antigen recognition have been previously reported. However, few studies focused on the meaning of harboring two residue types in position 37 of V H H antibodies, and the concrete roles of Y37 have been little to be elucidated. Here, we investigated the functional states of position 37 in co-crystal structures and performed analyses of three model antibodies with either F or Y at position 37. Our analysis indicates that Y at position 37 enhances the dissociation rate, which is highly correlated with drug efficacy. Our findings help to explain the molecular mechanisms that distinguish V H H antibodies from conventional antibodies., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier Inc.)

Several internal and external factors can influence animals' hormonal activity. Cortisol level in hair and wool determines chronic stress, which is connected with the long-term HPA axis effect. Wool cortisol levels in alpacas have never been determined to this time. The study aimed to assess the influence of selected factors on wool cortisol concentration in alpacas. The study included 36 alpacas. Wool samples were collected during shearing in June 2021, cut with an electric clipper from the right shoulder and the rump. Wool samples were fragmented into proximal (winter-spring regrowth) and distal (summer-fall regrowth) segments. Alpacas' Heat Stress Index (HSI) for the summer of 2020 was 139.4, and 116 for the winter of 2021. The cortisol levels in the wool samples were determined with the General Cortisol ELISA Kit assay. The most significant differences in wool cortisol concentrations were caused by two factors: the wool segment (P < 0.001; η2 = 0.889) and the region on the body (P < 0.001; η2 = 0.876). Wool cortisol level was higher in the distal segment (referring to the summer-fall season) than in the proximal one (referring to the winter-spring season). It is suggested that alpacas can feel heat stress in summer (HSI = 139.4), which could influence higher cortisol levels in the distal segment. The wool cortisol level was higher in the rump samples than the shoulder ones. Therefore, it is essential in future studies that wool samples from all tested animals should be completed from the same body region. Differences among age and sex groups were also observed. Wool cortisol level was higher in older animals, as differences between age groups were observed in samples from the rump in the distal and proximal segments (distal, the rump younger*older: P < 0.001; η 2  = 0.321; proximal, the rump older*younger: P = 0.007; η 2  = 0.195). Males showed higher cortisol levels than females, as a difference between sexes was observed in samples from the rump in the proximal segment (P = 0.001, η 2  = 0.271). This study emphasizes that various factors may significantly influence wool cortisol levels, which can be helpful in alpacas' welfare estimation using this hormonal indicator as a noninvasive long-term stress assessment method., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Alkaliphilic proteins have great potential as biocatalysts in biotechnology, especially for enzyme engineering. Extensive research has focused on exploring the enzymatic potential of alkaliphiles and characterizing alkaliphilic proteins. However, the current method employed for identifying these proteins that requires web lab experiment is time-consuming, labor-intensive, and expensive. Therefore, the development of a computational method for alkaliphilic protein identification would be invaluable for protein engineering and design. In this study, we present a novel approach that uses embeddings from a protein language model called ESM-2(3B) in a deep learning framework to classify alkaliphilic and non-alkaliphilic proteins. To our knowledge, this is the first attempt to employ embeddings from a pre-trained protein language model to classify alkaliphilic protein. A reliable dataset comprising 1,002 alkaliphilic and 1,866 non-alkaliphilic proteins was constructed for training and testing the proposed model. The proposed model, dubbed ALPACA, achieves performance scores of 0.88, 0.84, and 0.75 for accuracy, f1-score, and Matthew correlation coefficient respectively on independent dataset. ALPACA is likely to serve as a valuable resource for exploring protein alkalinity and its role in protein design and engineering., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Gastrointestinal adenocarcinomas are often reported in South American camelids (SAC). We describe here cases of gastroduodenal adenocarcinoma in an adult alpaca ( Vicugna pacos ) and a llama ( Llama glama ); both SACs were anorectic and lethargic before death. At autopsy, a prominent and firm caudal C3-pyloric-duodenal junction with stricture and ulceration was present in both animals, as were hemorrhages in various organs and hydrothorax. Microscopically, scattered nests, cords, and tortuous acini of neoplastic epithelial cells were embedded in desmoplastic stroma and invaded the submucosa and muscle layers of the gastroduodenal junction. The mucosa was necrotic, with gram-negative rods in the alpaca and colonies of gram-positive cocci in the llama. No tumor metastases were observed. The neoplastic cells immunolabeled for pancytokeratin. Escherichia coli was isolated from the alpaca and Streptococcus lutetiensis from the llama; septicemia was the cause of death in both animals. Although adenocarcinomas arising from gastric compartments and intestinal segments have been reported in SACs, adenocarcinoma of the caudal C3-pyloric-duodenal junction has not been reported previously in these species, to our knowledge., Competing Interests: Declaration of conflicting interestsThe authors declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.

Objective: To examine the prevalence of nasolacrimal duct (NLD) obstruction in hospital populations and assess signalment, diagnostics, and clinical approaches for alpacas and llamas diagnosed with NLD atresia or obstruction., Animals: 29 alpacas and 2 llamas., Clinical Presentation: Medical records were reviewed to identify camelids with NLD evaluated between 2000 and 2023. Signalment, history, physical examination data, diagnosis, and treatments were recorded. Follow-up information was gathered via phone and email questionnaire. Data were evaluated to determine prevalence and describe signalment and treatments for NLD disease., Results: 31 camelids met the inclusion criteria. A total of 8,027 alpacas and 1,998 llamas were seen within the study period at 1 teaching institution. The prevalence of NLD obstruction for this population of alpacas was 0.3% (26/8,027). Nineteen of 31 (61%) camelids presented at 1 year of age or younger. The most common physical exam finding was ocular discharge (68%). The most common diagnosis was NLD atresia (16/31 [51%]). Patency was established by surgical opening or lavage of the NLD. Long-term follow-up was available for 13 alpacas and 2 llamas, of which 11 (73%) had successful outcomes., Clinical Relevance: Results indicate that NLD obstruction is a condition that most commonly affects alpacas < 1 year of age and is associated with a good prognosis for treatment success.

Alpacas (Vicugna pacos) are reported to be the rare mammal in which the penis enters the uterus in mating. To date, however, only circumstantial evidence supports this assertion. Using female alpacas culled for meat, we determined that the alpaca penis penetrates to the very tips of the uterine horns, abrading the tract and breaking fine blood vessels. All female alpacas sacrificed one hour or 24 hours after mating showed conspicuous bleeding in the epithelium of some region of their reproductive tract, including the hymen, cervix and the tips of each uterine horn, but typically not in the vagina. Unmated females showed no evidence of conspicuous bleeding. Histological examination of mated females revealed widespread abrasion of the cervical and endometrial epithelium, injuries absent in unmated females. Within one hour of mating, sperm were already present in the oviduct. The male alpaca's cartilaginous penis tip with a hardened urethral process is likely responsible for the copulatory abrasion. The entire female reproductive tract interacts with the penis, functioning like a vagina. Alpacas are induced ovulators, and wounding may hasten delivery of the seminal ovulation-inducing factor beta-NGF into the female's blood stream. There is no evidence of sexual conflict in copulation in alpaca, and thus wounding may also be one of a variety of mechanisms devised by mammals to induce a beneficial, short-term inflammatory response that stimulates blastocyst implantation, the uterine remodeling associated with placental development, and thus the success of early pregnancy., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Brennan et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Background: Chimeric antigen receptor T (CAR-T) cell therapy, as an emerging anti-tumor treatment, has garnered extensive attention in the study of targeted therapy of multiple tumor-associated antigens in hepatocellular carcinoma (HCC). However, the suppressive microenvironment and individual heterogeneity results in downregulation of these antigens in certain patients' cancer cells. Therefore, optimizing CAR-T cell therapy for HCC is imperative., Methods: In this study, we administered FGFR4-ferritin (FGFR4-HPF) nanoparticles to the alpaca and constructed a phage library of nanobodies (Nbs) derived from alpaca, following which we screened for Nbs targeting FGFR4. Then, we conducted the functional validation of Nbs. Furthermore, we developed Nb-derived CAR-T cells and evaluated their anti-tumor ability against HCC through in vitro and in vivo validation., Results: Our findings demonstrated that we successfully obtained high specificity and high affinity Nbs targeting FGFR4 after screening. And the specificity of Nbs targeting FGFR4 was markedly superior to their binding to other members of the FGFR family proteins. Furthermore, the Nb-derived CAR-T cells, targeting FGFR4, exhibited significantly enhanced anti-tumor efficacy in both experiments when in vitro and in vivo., Conclusions: In summary, the results of this study suggest that the CAR-T cells derived from high specificity and high affinity Nbs, targeting FGFR4, exhibited significantly enhanced anti-tumor efficacy in vitro and in vivo. This is an exploration of FGFR4 in the field of Nb-derived CAR-T cell therapy for HCC, holding promise for enhancing safety and effectiveness in the clinical treatment of HCC in the future., (© 2024. The Author(s).)

Low motility and low sperm concentration are characteristics of alpaca semen. Thus, the intracytoplasmic sperm injection (ICSI) technique represents an alternative to improve the reproductive capacity of the male. However, the effect of post-ICSI activation in alpaca is not yet known. The aim of the present study was to compare the effect of chemical activators on alpaca embryo development after ICSI. Alpaca ovaries were collected from a local slaughterhouse and transported to the laboratory. Category I, II and III oocytes were matured for 30 h at 38.5 °C. After ICSI, injected oocytes were randomly divided and activated as follows: i) 5 μM ionomycin for 5 min, ii) 7% ethanol for 4 min, iii) 5 μM ionomycin for 5 min, window period 3 h plus 7% ethanol for 4 min, iv) 5 μM ionomycin for 5 min, window period 3 h, a second ionomycin treatment for 5 min, followed by 1.9 mM 6-DMAP for 3 h, v) 10 mM SrCl 2 for 3 h. Culture was carried out for 5 days in SOFaa at 38.5 °C. The cleavage rate was the lowest in the SrCl 2 group, morula development was the lowest in the SrCl 2 and without activation groups, and blastocyst stage was not different between groups (P<0.05). The rates with SrCl 2 were lower in total embryos produced, whereas in transferable embryos they were lower with 2Io/6-DMAP and with SrCl 2 (P<0.05). In conclusion, alpaca oocyte activation is more efficient with ionomycin and ethanol to produce transferable embryos., Competing Interests: Declaration of Competing Interest The authors declare no potential conflicts of interest., (Copyright © 2024 Elsevier B.V. All rights reserved.)

South American camelids (SACs) play an increasing role in veterinary care in Europe. Many alpacas or llamas presented to veterinarians suffer from anaemia, regularly with a packed cell volume (PCV) below 0.10 l/l, which is a life-threatening condition for the animals. This review article presents clinical and laboratory diagnostic tools for the diagnosis of anaemia in SACs. Clinical identification of anaemic animals can be performed by assessing the FAMACHA© score and the Body Condition Score (BCS), since anaemia in alpacas and llamas correlates with pale mucous membranes and a lowered BCS. Haematological examination of a blood sample can provide a more differentiated diagnosis of anaemia in SACs. A common finding is regenerative anaemia with an increased number of reticulocytes that is often caused by blood loss due to Haemonchus contortus. Changes in a blood smear from an alpaca or llama with regenerative anaemia may include normoblasts (nucleated red blood cells), anisocytosis, poikilocytosis, polychromasia, Howell-Jolly bodies or basophilic stippling. Furthermore, non-regenerative anaemia, often caused by trace element deficiency or cachexia, can also occur., (© 2023. The Author(s).)

Assessment of the body condition score (BCS) is an important tool to check the nutritional status of an animal, and thus can provide important information in herd management, but also in the veterinary examination of individual animals. In dairy farming, BCS has been recognized as an important parameter for a long time already, but over the years body condition scoring has also become more important for other species. Especially in South American camelids (SACs: alpacas and llamas), whose dense coat can hide a poor nutritional status for a long time, regular assessment of the BCS is recommended by different authors and organizations. To date, there are several instructions for the assessment of the BCS in SACs, which, however, differ significantly in some points. For a consistent approach, a total of 35 instructions for the BCS in SACs were compared in a scoping review. The spine, especially the lumbar vertebrae as well as the ribs were identified as the two main body sites recommended for assessing the BCS. Furthermore, the area between the front and rear leg and the pelvis were mentioned frequently; however, these body sites were discussed controversially. The paralumbar fossa, shoulder, and udder were only mentioned sporadically. Among the various scales, the 1-5 scale was mentioned most frequently, with the optimal BCS being 3. Nonetheless, physiological fluctuation due to breed, age, sex, pregnancy, and lactation have to be considered. The BCS should be assessed each time the animals are handled, but at least monthly. In contrast to body weight, the BCS reflects the nutritional status of an animal even after a single examination. The review showed that much of the available information on the assessment of BCS in SACs is based on practical experience rather than scientific evidence. Indeed, some of the assumptions made there might still have to be verified or modified., (© 2023. The Author(s).)

The expression level of PD-L1 in tumor tissue is considered one of the effective biomarkers to guide PD-1/PD-L1 therapy. Quantifying whole-body PD-L1 expression by SPECT imaging may help in selecting patients that potentially respond to PD-1/PD-L1 therapy. Nanobody is the smallest antibody fragment with antigen-binding ability that is well suited for radionuclide imaging. Nevertheless, high retention of radioactivity in the kidney may limit its clinical translation. The present study aimed to screen, design, and prepare a nanobody-based SPECT probe with rapid renal clearance to evaluate the PD-L1 expression level in vivo noninvasively. A phage library was constructed by immunizing alpaca with recombinant human PD-L1 protein, and 17 anti-PD-L1 nanobodies were screened by the phage display technique. After sequence alignment and flow cytometry analysis, APN09 was selected as the candidate nanobody, and a GGGC chelator was attached to its C-terminus for 99m Tc labeling to prepare a SPECT imaging probe. The affinity and specificity of 99m Tc-APN09 were evaluated by protein and cell-binding experiments, and SPECT imaging and biodistribution were performed in a mouse model with bilateral transplantation of A549 and A549 PD-L1 tumors. The ability of 99m Tc-APN09 to quantify the PD-L1 expression level in vivo was validated in tumor models with different PD-L1 expression levels. 99m Tc-APN09 had a radiochemical purity higher than 99% and a binding equilibrium dissociation constant of 21.44 ± 1.65 nM with hPD-L1, showing high affinity. SPECT imaging results showed that 99m Tc-APN09 could efficiently detect PD-L1-positive tumors within 0.5 h, and the quantitative results of SPECT were well correlated with the expression level of PD-L1 in cell lines. SPECT imaging and biodistribution results also showed that 99m Tc-APN09 was rapidly cleared from the kidney in 2 h postinjection. 99m Tc-APN09 was a simple and stable tool for visualizing PD-L1 expression in the whole body. In addition, due to its significant reduction in renal retention, it has better prospects for clinical translation.

In the last years, there has been an increasing interest in llamas, not only as part of a productive system, but mostly as companion animals. Most reports regarding clinical biochemistry and haematology include few llamas and details about their health status are not available. The present study aims to provide haematological and biochemical parameters for llamas of known health status. Twenty-three non-pregnant females and seven males that live in Buenos Aires, Argentina (34°36'S, 58°22'W, at sea level) were studied. Llamas were clinically healthy, in good nutritional status. Animals were kept at grass and were fed hay bale or pellets and water ad libitum. Blood samples were collected by jugular venipuncture in spring. Packed cell volume, leucocyte count, differential white cell count, platelets count, urea, creatinine, total proteins, albumin, alkaline phosphatase, aspartate aminotransferase, alanine aminotransferase, glucose, calcium and phosphate were assessed. No significant differences were observed between males and females, except for platelet count and calcium, which was greater in males (P˂0.01). Values obtained for the different parameters were similar to those previously reported, except for monocytes, alkaline phosphatase, glucose and calcium, that were lower and lymphocytes and platelets count, that were higher in this study. In conclusion, different ambient and methodological conditions might affect some parameters. The parameters hereby presented are representative of llama's population living at sea level in South America., (© 2023. The Author(s), under exclusive licence to Springer Nature B.V.)

Motivation: Large Language Models (LLMs) have the potential to revolutionize the field of Natural Language Processing, excelling not only in text generation and reasoning tasks but also in their ability for zero/few-shot learning, swiftly adapting to new tasks with minimal fine-tuning. LLMs have also demonstrated great promise in biomedical and healthcare applications. However, when it comes to Named Entity Recognition (NER), particularly within the biomedical domain, LLMs fall short of the effectiveness exhibited by fine-tuned domain-specific models. One key reason is that NER is typically conceptualized as a sequence labeling task, whereas LLMs are optimized for text generation and reasoning tasks., Results: We developed an instruction-based learning paradigm that transforms biomedical NER from a sequence labeling task into a generation task. This paradigm is end-to-end and streamlines the training and evaluation process by automatically repurposing pre-existing biomedical NER datasets. We further developed BioNER-LLaMA using the proposed paradigm with LLaMA-7B as the foundational LLM. We conducted extensive testing on BioNER-LLaMA across three widely recognized biomedical NER datasets, consisting of entities related to diseases, chemicals, and genes. The results revealed that BioNER-LLaMA consistently achieved higher F1-scores ranging from 5% to 30% compared to the few-shot learning capabilities of GPT-4 on datasets with different biomedical entities. We show that a general-domain LLM can match the performance of rigorously fine-tuned PubMedBERT models and PMC-LLaMA, biomedical-specific language model. Our findings underscore the potential of our proposed paradigm in developing general-domain LLMs that can rival SOTA performances in multi-task, multi-domain scenarios in biomedical and health applications., Availability and Implementation: Datasets and other resources are available at https://github.com/BIDS-Xu-Lab/BioNER-LLaMA., (© The Author(s) 2024. Published by Oxford University Press.)

Objectives: To evaluate the effectiveness of safeguards to prevent large language models (LLMs) from being misused to generate health disinformation, and to evaluate the transparency of artificial intelligence (AI) developers regarding their risk mitigation processes against observed vulnerabilities., Design: Repeated cross sectional analysis., Setting: Publicly accessible LLMs., Methods: In a repeated cross sectional analysis, four LLMs (via chatbots/assistant interfaces) were evaluated: OpenAI's GPT-4 (via ChatGPT and Microsoft's Copilot), Google's PaLM 2 and newly released Gemini Pro (via Bard), Anthropic's Claude 2 (via Poe), and Meta's Llama 2 (via HuggingChat). In September 2023, these LLMs were prompted to generate health disinformation on two topics: sunscreen as a cause of skin cancer and the alkaline diet as a cancer cure. Jailbreaking techniques (ie, attempts to bypass safeguards) were evaluated if required. For LLMs with observed safeguarding vulnerabilities, the processes for reporting outputs of concern were audited. 12 weeks after initial investigations, the disinformation generation capabilities of the LLMs were re-evaluated to assess any subsequent improvements in safeguards., Main Outcome Measures: The main outcome measures were whether safeguards prevented the generation of health disinformation, and the transparency of risk mitigation processes against health disinformation., Results: Claude 2 (via Poe) declined 130 prompts submitted across the two study timepoints requesting the generation of content claiming that sunscreen causes skin cancer or that the alkaline diet is a cure for cancer, even with jailbreaking attempts. GPT-4 (via Copilot) initially refused to generate health disinformation, even with jailbreaking attempts-although this was not the case at 12 weeks. In contrast, GPT-4 (via ChatGPT), PaLM 2/Gemini Pro (via Bard), and Llama 2 (via HuggingChat) consistently generated health disinformation blogs. In September 2023 evaluations, these LLMs facilitated the generation of 113 unique cancer disinformation blogs, totalling more than 40 000 words, without requiring jailbreaking attempts. The refusal rate across the evaluation timepoints for these LLMs was only 5% (7 of 150), and as prompted the LLM generated blogs incorporated attention grabbing titles, authentic looking (fake or fictional) references, fabricated testimonials from patients and clinicians, and they targeted diverse demographic groups. Although each LLM evaluated had mechanisms to report observed outputs of concern, the developers did not respond when observations of vulnerabilities were reported., Conclusions: This study found that although effective safeguards are feasible to prevent LLMs from being misused to generate health disinformation, they were inconsistently implemented. Furthermore, effective processes for reporting safeguard problems were lacking. Enhanced regulation, transparency, and routine auditing are required to help prevent LLMs from contributing to the mass generation of health disinformation., Competing Interests: Competing interests: All authors have completed the ICMJE uniform disclosure form at www.icmje.org/disclosure-of-interest/ and declare: AMH holds an emerging leader investigator fellowship from the National Health and Medical Research Council (NHMRC), Australia; NDM is supported by a NHMRC postgraduate scholarship, Australia; MJS is supported by a Beat Cancer research fellowship from the Cancer Council South Australia; BDM’s PhD scholarship is supported by The Beat Cancer Project, Cancer Council South Australia, and the NHMRC, Australia; no support from any other organisation for the submitted work; AR and MJS are recipients of investigator initiated funding for research outside the scope of the current study from AstraZeneca, Boehringer Ingelheim, Pfizer, and Takeda; and AR is a recipient of speaker fees from Boehringer Ingelheim and Genentech outside the scope of the current study. There are no financial relationships with any other organisations that might have an interest in the submitted work in the previous three years to declare; no other relationships or activities that could appear to have influenced the submitted work., (© Author(s) (or their employer(s)) 2019. Re-use permitted under CC BY-NC. No commercial re-use. See rights and permissions. Published by BMJ.)

It is likely that individuals are turning to Large Language Models (LLMs) to seek health advice, much like searching for diagnoses on Google. We evaluate clinical accuracy of GPT-3·5 and GPT-4 for suggesting initial diagnosis, examination steps and treatment of 110 medical cases across diverse clinical disciplines. Moreover, two model configurations of the Llama 2 open source LLMs are assessed in a sub-study. For benchmarking the diagnostic task, we conduct a naïve Google search for comparison. Overall, GPT-4 performed best with superior performances over GPT-3·5 considering diagnosis and examination and superior performance over Google for diagnosis. Except for treatment, better performance on frequent vs rare diseases is evident for all three approaches. The sub-study indicates slightly lower performances for Llama models. In conclusion, the commercial LLMs show growing potential for medical question answering in two successive major releases. However, some weaknesses underscore the need for robust and regulated AI models in health care. Open source LLMs can be a viable option to address specific needs regarding data privacy and transparency of training., (© 2024. The Author(s).)

Large language models (LLMs) have the potential to transform our lives and work through the content they generate, known as AI-Generated Content (AIGC). To harness this transformation, we need to understand the limitations of LLMs. Here, we investigate the bias of AIGC produced by seven representative LLMs, including ChatGPT and LLaMA. We collect news articles from The New York Times and Reuters, both known for their dedication to provide unbiased news. We then apply each examined LLM to generate news content with headlines of these news articles as prompts, and evaluate the gender and racial biases of the AIGC produced by the LLM by comparing the AIGC and the original news articles. We further analyze the gender bias of each LLM under biased prompts by adding gender-biased messages to prompts constructed from these news headlines. Our study reveals that the AIGC produced by each examined LLM demonstrates substantial gender and racial biases. Moreover, the AIGC generated by each LLM exhibits notable discrimination against females and individuals of the Black race. Among the LLMs, the AIGC generated by ChatGPT demonstrates the lowest level of bias, and ChatGPT is the sole model capable of declining content generation when provided with biased prompts., (© 2024. The Author(s).)

Background: The global spread of extended-spectrum β-lactamase (ESβL)-producing Escherichia coli has been considered a One Health issue that demands continuous genomic epidemiology surveillance in humans and non-human hosts., Objectives: To report the occurrence and genomic data of ESβL-producing E. coli strains isolated from South American llamas inhabiting a protected area with public access in the Andean Highlands of Peru., Methods: Two ESβL-producing E. coli strains (E. coli L1LB and L2BHI) were identified by MALDI-TOF. Genomic DNAs were extracted and sequenced using the Illumina NextSeq platform. De novo assembly was performed by CLC Genomic Workbench and in silico prediction was accomplished by curated bioinformatics tools. SNP-based phylogenomic analysis was performed using publicly available genomes of global E. coli ST10., Results: Escherichia coli L1LB generated a total of 4 000 11 and L2BHI a total of 4 002 54 paired-end reads of ca.164 × and ca. 157 ×, respectively. Both E. coli strains were assigned to serotype O8:H4, fimH41, and ST10. The bla CTX-M-65 ESβL gene, along with other medically important antimicrobial resistance genes, was predicted. Broad virulomes, including the presence of the astA gene, were confirmed. The phylogenomic analysis revealed that E. coli L1LB and L2BHI strains are closely related to isolates from companion animals and human hosts, as well as environmental strains, previously reported in North America, South America, Africa, and Asia., Conclusion: Presence of ESβL-producing E. coli ST10 in South American camelids with historical and cultural importance supports successful expansion of international clones of priority pathogens in natural areas with public access., (Copyright © 2023. Published by Elsevier Ltd.)

This study evaluated the ability of generative large language models (LLMs) to detect speech recognition errors in radiology reports. A dataset of 3233 CT and MRI reports was assessed by radiologists for speech recognition errors. Errors were categorized as clinically significant or not clinically significant. Performances of five generative LLMs-GPT-3.5-turbo, GPT-4, text-davinci-003, Llama-v2-70B-chat, and Bard-were compared in detecting these errors, using manual error detection as the reference standard. Prompt engineering was used to optimize model performance. GPT-4 demonstrated high accuracy in detecting clinically significant errors (precision, 76.9%; recall, 100%; F1 score, 86.9%) and not clinically significant errors (precision, 93.9%; recall, 94.7%; F1 score, 94.3%). Text-davinci-003 achieved F1 scores of 72% and 46.6% for clinically significant and not clinically significant errors, respectively. GPT-3.5-turbo obtained 59.1% and 32.2% F1 scores, while Llama-v2-70B-chat scored 72.8% and 47.7%. Bard showed the lowest accuracy, with F1 scores of 47.5% and 20.9%. GPT-4 effectively identified challenging errors of nonsense phrases and internally inconsistent statements. Longer reports, resident dictation, and overnight shifts were associated with higher error rates. In conclusion, advanced generative LLMs show potential for automatic detection of speech recognition errors in radiology reports. Keywords: CT, Large Language Model, Machine Learning, MRI, Natural Language Processing, Radiology Reports, Speech, Unsupervised Learning Supplemental material is available for this article ., (© RSNA, 2024.)

Large animals could be important drivers of spatial nutrient subsidies when they ingest resources in some habitats and release them in others, even moving nutrients against elevational gradients. In high Andean deserts, vicuñas (Vicugna vicugna) move daily between nutrient-rich wet meadows, where there is abundant water and forage but high risk of predation by pumas (Puma concolor), and nutrient-poor open plains with lower risk of predation. In all habitats, vicuñas defecate and urinate in communal latrines. We investigated how these latrines impacted soil and plant nutrient concentrations across three habitats in the Andean ecosystem (meadows, plains, and canyons) and used stable isotope analysis to explore the source of fecal nutrients in latrines. Latrine soils had higher concentrations of nitrogen, carbon, and other nutrients than did nonlatrine soils across all habitats. These inputs corresponded with an increase in plant quality (lower C:N) at latrine sites in plains and canyons, but not in meadows. Stable isotope mixing models suggest that ~7% of nutrients in plains latrines originated from vegetation in meadows, which is disproportionately higher than the relative proportion of meadow habitat (2.6%) in the study area. In contrast, ~68% of nutrients in meadow latrines appear to originate from plains and canyon vegetation, though these habitats made up nearly 98% of the study area. Vicuña diel movements thus appear to concentrate nutrients in latrines within habitats and to drive cross-habitat nutrient subsidies, with disproportionate transport from low-lying, nutrient-rich meadows to more elevated, nutrient-poor plains. When these results are scaled up to the landscape scale, the amount of nitrogen and phosphorus subsidized in soil at plains latrines was of the same order of magnitude as estimates of annual atmospheric nitrogen and phosphorus deposition for this region (albeit far more localized and patchy). Thus, vicuña-mediated nutrient redistribution and deposition appears to be an important process impacting ecosystem functioning in arid Andean environments, on par with other major inputs of nutrients to the system., (© 2024 The Ecological Society of America.)