Castration Research Articles

Exploring the criteria influencing construction productivity of diaphragm walls in underground stations in Egypt

Planning and cost estimation engineers face great challenges to estimate construction productivity rate (PR) of diaphragm walls (DWs). A specific guideline to predict the construction of DWs in underground station is still not available. In order to overcome these challenges, the criteria affecting PR for the construction of DWs should be hence defined and the factors that impact the construction productivity of DWs should be well understood. Briefly and given the limitations of previous research work, we gathered and compiled a comprehensive key list of all criteria based on previous reports. This comprehensive key list was later reviewed by underground construction and specialized experts to define, identify and distinguish the most important criterion affecting the construction productivity of DWs in Egypt. We developed two questionnaire surveys to conduct these interviews with the experts. This obtained criterion was ranked and weighted using Simos’ procedure based on prioritizing the influencing criteria. Our findings concluded that the ground condition type/and characteristics, soil–machine interaction, machine type and model characteristics (grab/or cutter), breakdowns and de-sender efficiency are significantly influencing the excavation rate. Additionally, factors such as cage type (fiber/or steel), connection type (welding/or mechanical) and volume of steel fixing crew are crucial to identify the rate of installation cages. On the other side, the arrival time of concrete car mixer and panel volume were determined to significantly impact the concrete casting rate. In summary, this study presented a distinctive weighting and ranking methodology of the influencing criteria for the PR of DWs. Furthermore, this work provides an initial concept for many full-scale prediction applications to estimate an accurate construction productivity of DWs, particularly in the underground projects.

Castration for osteomalacia

As the essence of osteomalacia, so even more the possibility of its cure by castration represent many mysterious and unexplained sides [cf. Jury. obstetrician. and women. Bol. , 1888, p. 938 and 1890, p. 837. Ref ..] In view of this, every observation related to this is extremely important.

Testosterone deficiency caused by castration increases adiposity in male rats in a tissue-specific and diet-dependent manner

BackgroundTestosterone deficiency in men is clinically associated with the development of metabolic syndrome, which manifests as obesity, hepatic steatosis, and type-2 diabetes. We investigated the effects of castration-induced testosterone deficiency on body adiposity and the expression of genes related to lipid metabolism and glucose uptake and androgen signaling in male rats fed a normal diet (ND) or a high-fat diet (HFD).MethodsChanges in lipid and glucose metabolism and androgen signaling were investigated at physiological and molecular levels in the muscle, liver, and adipose tissues of non-castrated and castrated rats under ND or HFD feeding.ResultsCastration-induced testosterone deficiency predisposed animals on ND to early development of fatty liver by activating fatty acid (FA) synthesis, whereas HFD activated hepatic FA uptake CD36 expression, leading to the development of hepatic steatosis. In rats fed ND, castration induced muscle fat accumulation by activating CD36 expression. In the subcutaneous fat of ND-fed rats, castration increased adiposity and the expression of FA synthesis-related genes, but it decreased glucose transporter gene expression. In the abdominal fat of rats fed ND, castration increased adiposity by upregulating FA synthesis-related genes, and HFD promoted adiposity by inducing FA uptake, glucose transporter, and FA synthesis-related gene expression. In rats fed ND, castration decreased body growth and muscle weight and downregulated the expression of genes androgen signaling in the longissimus dorsi muscle.ConclusionsTestosterone deficiency increases adiposity in a tissue-specific and diet-dependent manner. Testosterone deficiency decreases body and muscle weights and downregulates androgen signaling.

Abstract 381: Macrophage tracking using multi-modality 3D imaging in xenografts

Abstract A common approach to immunotherapy is to promote the accumulation of specific bone marrow-derived myeloid cells into tumors for anti-tumor activity. While the pro- and anti-tumor effects of differentiated myeloid cells are complex, several novel therapeutic approaches are focused on recruiting macrophages to tumors and transforming their pro-tumor phenotypes of macrophages to anti-tumor phenotypes. Research efforts to better understand and manipulate this type of macrophage biology is critical for developing and testing therapeutics. These experimental models commonly utilize imaging approaches, be it in vivo using molecular imaging or ex vivo histological approaches, to visualize macrophages in the context of tumors. These different imaging modalities provide different advantages in their ability characterize macrophage distribution, tissue localization and kinetics: In vivo imaging, such as MRI and in vivo fluorescence, gives strong temporal insights and general distribution in target tissues but are limited by sensitivity and resolution. Fluorescence microscopy and histology provide substantially higher resolution and sensitivity regarding TAM localization and cellular morphology but lacks temporal resolution and is only sampling a small portion of target tissue. Cryofluorescence tomography (CFT) is an attractive new modality in which an entire animal or tissue can be imaged ex vivo in 3D with high-resolution and increased sensitivity compared to nuclear in vivo imaging modalities. To harness the advantages of each imaging modality, we designed a multimodality imaging study enabling the utilization of MRI, FLI and CFT for in vivo and ex vivo visualization of macrophages in tumor bearing animals. In this study, mice bearing xenografts were administered with V-Sense (VS-1000H NIR, Celsense, Inc., Pittsburgh, PA), a perfluorocarbon emulsion containing fluorine-19 (19F) and a NIR fluorophore. When administered IV, V-Sense is preferentially taken up by cells of the recticuloendothlium system, including Kupffer cells and macrophages, especially in inflamed tissues (e.g., tumors) thus enabling imaging using both MRI and fluorescence imaging. Immediately following MRI, animals underwent FLI and were then frozen for CFT. As expected, 19F signal was evident in mice whose tumors where acute inflammation was present, ) but not detected in the non-inflamed tumors, indicative for macrophage recruitment. However, when imaged using both FLI and CFT, V-Sense signal was detected in tumors of both groups (1 and 2). CFT proved to be the most sensitive, revealing V-Sense signal not only in the tumor and liver but also in several other organs such as lymph nodes and bone marrow that were undetected with MRI or FLI. The results of this study demonstrated the added value of a multimodality imaging approach using V-Sense, in which low resolution in vivo imaging and high-resolution 3D ex vivo CFT imaging can complement each other, providing rich, multi-resolution layers of immunological information in the same subjects. Citation Format: Hemi Dimant, Patrick McConville, Kelly Orcutt, Kieth Mikule, Keith Wilcoxen, Deanne Lister, Eric T. Ahrens. Macrophage tracking using multi-modality 3D imaging in xenografts [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 381.

Risk or Protective Factors of Prostate Cancer Progressing to Castration Resistant Prostate Cancer After Androgen Deprivation Therapy

Risk or Protective Factors of Prostate Cancer Progressing to Castration Resistant Prostate Cancer After Androgen Deprivation Therapy

Involvement of sex hormones, oxidative stress, ACE and ACE2 activity in the impairment of renal function and remodelling in SHR

AimsHypertension is a relevant sex and sex hormones-dependent risk factor where the cardiovascular and renal health of the population are concerned. Men experience greater losses of renal function (RF) than women, but the mechanisms remain somewhat unclear. Our goal was to evaluate the relationship between oxidative stress (OS), angiotensin-converting enzyme (ACE) and angiotensin-converting enzyme 2 (ACE2) activities and RF in male and female SHR. Main methodsTwelve-week-old spontaneously hypertensive rats (SHR) were submitted to either castration or SHAM surgery and divided into 4 groups, SHAM or Castrated (CAST) males or females. After 51 days we evaluated RF (inulin and sodium para-aminohippurate), ACE and ACE2 activities (fluorimetry), OS (flow cytometry), collagen deposition (picrosirius red) and protein expression (western blot). Key findingsMales presented lower RF than females and castration impaired this parameter in both groups. Sexual dimorphism was not observed regarding OS and inflammation; however, castration increased this parameter more severely in males than in females. SHAM males exhibited higher collagen deposition than females, though castration increased it in both sexes, eliminating the difference. We found sexual dimorphism regarding renal ACE and ACE2 activities, which were lower in males than in females. Although castration did not alter ACE activity, it reduced ACE2 activity in females and increased it in males. SignificanceThese results indicate that sex hormones affect RF in SHR. As alterations in the oxidative system were capable of promoting podocyte injury, inflammation, and collagen deposition, we put forward that these effects are differently modulated by ACE and ACE2.

Use of masking strategies to avoid the boar taint perception in chorizo: consumers' acceptability

The production of entire male pigs is an alternative to surgical castration. However, boar taint may be present in the meat of entire male animals. The aim of the present paper was to investigate the acceptability and sensory properties of chorizo, elaborated under three different levels of androstenone [castrated 0–0.3 mg / kg, medium 0.4–0.9 mg / kg and high 1–2.75 mg / kg] and two masking strategies [rosemary, and rosemary plus smoking] using a Check-All-That-Apply questionnaire. The results showed that, of the strategies employed, the rosemary plus smoked chorizo was the most effective masking strategy to reduce the boar taint in chorizos with high androstenone level (3.7% selection rate for animal flavour), while the closest to the ideal product, according to consumers' description, was high androstenone level plus rosemary chorizo.

The connectome of the Caenorhabditis elegans pharynx.

Detailed anatomical maps of individual organs and entire animals have served as invaluable entry points for ensuing dissection of their evolution, development, and function. The pharynx of the nematode Caenorhabditis elegans is a simple neuromuscular organ with a self-contained, autonomously acting nervous system, composed of 20 neurons that fall into 14 anatomically distinct types. Using serial electron micrograph (EM) reconstruction, we re-evaluate here the connectome of the pharyngeal nervous system, providing a novel and more detailed view of its structure and predicted function. Contrasting the previous classification of pharyngeal neurons into distinct inter- and motor neuron classes, we provide evidence that most pharyngeal neurons are also likely sensory neurons and most, if not all, pharyngeal neurons also classify as motor neurons. Together with the extensive cross-connectivity among pharyngeal neurons, which is more widespread than previously realized, the sensory-motor characteristics of most neurons define a shallow network architecture of the pharyngeal connectome. Network analysis reveals that the patterns of neuronal connections are organized into putative computational modules that reflect the known functional domains of the pharynx. Compared with the somatic nervous system, pharyngeal neurons both physically associate with a larger fraction of their neighbors and create synapses with a greater proportion of their neighbors. We speculate that the overall architecture of the pharyngeal nervous system may be reminiscent of the architecture of ancestral, primitive nervous systems.

Inhibitory effect of ergosterol on bladder carcinogenesis is due to androgen signaling inhibition by brassicasterol, a metabolite of ergosterol.

We previously revealed that Choreito, a traditional Kampo medicine, strongly inhibits bladder carcinogenesis promotion. We have also shown that Polyporus sclerotium, which is one of the crude drugs in Choreito, has the strongest bladder carcinogenesis inhibitory effect and that the ergosterol contained in Polyporus sclerotium is the main active component. In this study, we analyzed the mechanism by which ergosterol inhibits bladder carcinogenesis. Rats were given an N-butyl-N-(4-hydroxybutyl) nitrosamine (BHBN) solution ad libitum, and then a promoter [saccharin sodium (SS), DL-tryptophan, or BHBN] was administered together with ergosterol or its metabolite, brassicasterol. The bladders were removed from rats, and the inhibitory effect on carcinogenesis promotion was evaluated by an agglutination assay with concanavalin A (Con A). Although the oral administration of ergosterol inhibited the promotion of bladder carcinogenesis with SS, the intraperitoneal administration of brassicasterol showed a stronger effect. The effect of brassicasterol on carcinogenesis promotion was observed regardless of the type of promoter. Administration of testosterone to castrated rats increased the number of cell aggregates caused by Con A. In contrast, intraperitoneal administration of brassicasterol to castrated rats treated with testosterone significantly decreased the number of cell aggregates, confirming the inhibition of bladder carcinogenesis promotion. The inhibitory effect of ergosterol on bladder carcinogenesis is due to brassicasterol, a metabolite of ergosterol. The action of brassicasterol via androgen signaling may play a role in the inhibitory effect on bladder carcinogenesis promotion.

Effect of Gonadectomy and Angiotensin II Receptor Blockade in a Mouse Model of Isoproterenol-induced Cardiac Diastolic Dysfunction.

Although heart failure (HF) with preserved ejection fraction (HFpEF) is more common in postmenopausal women than in men, the effect of sex hormones on cardiac diastolic function remains unclear. We examined the effect of gonadectomy with or without the angiotensin receptor blocker olmesartan (Olm) in an isoproterenol (ISO) -induced mouse model of left ventricular hypertrophy (LVH) and cardiac diastolic dysfunction. ISO or ISO with Olm were administered for 28 days in sham-operated male and female, castrated (CAS), and ovariectomized (OVX) mice. LV ejection fraction (EF) and E/A ratio were analyzed by echocardiography, and the LV and lung weight corrected by tibial length were used as indices of LVH and lung congestion, respectively. On echocardiography, systolic function did not differ between the four groups. LV/tibial length (TL) and Lung/TL significantly increased in all groups. The LV/TL ratio was lower in castrated-ISO vs. Male-Sham-ISO but did not differ between Female-Sham-ISO and OVX-ISO. However, the Lung/TL ratio of OVX-ISO was greater than that of Female-Sham-ISO. Olm prevented LV hypertrophy in all groups. The decrease in E/A and increase in lung weight were improved by Olm in Male-Sham and OVX-ISO but not in the other groups. These sex differences suggest that sex hormones play a pivotal role in modulating cardiac hypertrophy and diastolic dysfunction induced by chronic β-adrenoceptor stimulation, and thus affect the therapeutic potential of angiotensin receptor blockade.

Testosterone-androgen receptor: The steroid link inhibiting TRPM8-mediated cold sensitivity.

Recent studies have revealed gender differences in cold perception, and pointed to a possible direct action of testosterone (TST) on the cold-activated TRPM8 (Transient Receptor Potential Melastatin Member 8) channel. However, the mechanisms by which TST influences TRPM8-mediated sensory functions remain elusive. Here, we show that TST inhibits TRPM8-mediated mild-cold perception through the noncanonical engagement of the Androgen Receptor (AR). Castration of both male rats and mice increases sensitivity to mild cold, and this effect depends on the presence of intact TRPM8 and AR. TST in nanomolar concentrations suppresses whole-cell TRPM8-mediated currents and single-channel activity in native dorsal root ganglion (DRG) neurons and HEK293 cells co-expressing recombinant TRPM8 and AR, but not TRPM8 alone. AR cloned from rat DRGs shows no difference from standard AR. However, biochemical assays and confocal imaging reveal the presence of AR on the cell surface and its interaction with TRPM8 in response to TST, leading to an inhibition of channel activity.

Protocatechuic acid ameliorates testosterone‐induced benign prostatic hyperplasia through the regulation of inflammation and oxidative stress in castrated rats

Protocatechuic acid (PA) is a polyphenol-recognized for its efficacy as an antioxidant-possesses anticancer, anti-inflammatory, antioxidant properties. The efficacy of PA in the management of benign prostatic hyperplasia (BPH) has not been investigated. Forty-two castrated rats (n = 7) were treated as follows: control (corn oil), BPH only received testosterone propionate (TP) (TP 3 mg/kg intraperitoneally), BPH + PA (TP 3 mg/kg + PA 40 mg/kg), BPH + finasteride (Fin) (TP 3 mg/kg + Fin 10 mg/kg), PA only (40 mg/kg: by gavage), and Fin only (10 mg/kg: by gavage) for 4 weeks. In BPH rats, there were significant (P < .05) increases in prostatic (250%) and organosomatic (280%) weights compared with controls. Cotreatment decreased prostatic weights by 19% (PA) and 21% (Fin). Markers of inflammation: myeloperoxidase activities increased in serum (148%) and prostate (70%), as well as nitric oxide levels serum (92%) and prostatic (95%). Proinflammatory cytokines interleukin-1β and tumor necrosis factor-α increased by 3.6- and 2.8-fold. Furthermore, prostatic malondialdehyde, superoxide dismutase, and serum total acid phosphatase increased by 97%, 25%, and 48%, respectively. Histology revealed poor architecture and severe proliferation of the prostate in BPH rats. Inflammation and oxidative stress markers, as well as the histological alteration in BPH rats, was attenuated (P < .05) upon cotreatment with PA and comparable with Fin cotreatment. These results suggest that PA mitigates oxido-inflammatory responses and restored prostatic cytoarchitecture to levels comparable with control in rats induced with BPH.

Tumor-Associated Macrophages (TAM) are recruited to the aging prostate epithelial lesions and become intermingled with basal cells.

Prostate cancer remains one of the most common cancers in men. Macrophages are thought to be important regulators in cancers, and their potential involvement in prostate cancer should not be overlooked. Therefore, the association between macrophages and the pre-tumorous changes in prostate epithelium during aging deserves further investigation. We sought to investigate whether macrophages would be recruited into the prostate epithelium that display pathological lesions commonly found during aging. Prostates of aging rats, with and without treatment with a combination of testosterone and estradiol, were examined for premalignant and malignant epithelial lesions. For comparison, prostates of castrated rats were also investigated. Intraepithelial macrophages were found restricted to areas of premalignant and malignant lesions. An unprecedented interaction between macrophages and basal cells was observed in the aging pathological lesions. The intraepithelial macrophages were associated with autophagy, in contrast to those found after castration. In prostate lesions, the intraepithelial macrophages had TAM phenotype (CD68+/iNOS+/CD206+/ARG+), denoting a possible involvement in cancer progression. However, M2 macrophages (CD68+/CD163+) were recruited into the epithelium after castration, possibly to phagocytize cells undergoing apoptosis. In conclusion, macrophages were recruited into the prostate epithelium and presented diverse phenotypes and morphology, consistent with changes reflected in the hormonal environment. Macrophages with the TAM phenotype were found restricted to areas of premalignant and malignant lesions in aging prostates, denoting a possible involvement in cancer progression. In contrast, M2 macrophages were found in the regressed epithelium after castration.

The value of AR-V7 in predicting therapeutic efficiency of abiraterone for CRPC patients

Objective Determine the relationship between AR-V7 expression and treatment efficiency of abiraterone. Methods A prospective cohort study was conducted to detect CTCs AR-V7 in newly diagnosed CRPC patients, aged ≥18 years who were admitted to the urologic department of Peking University cancer hospital from January 2016 to January 2019, with one or more systemic metastases. All the patients intended to be treated with abiraterone. According to the AR-V7 status, patients were divided into 2 groups (AR-V7 positive and negative). PSA decline time, PSA PFS, clinical status PFS, imaging PFS and CSS are analyzed and compared by t-test and Chi-square between 2 groups. Result 49 patients were AR-V7 negative and 28 were AR-V7 positive. Compared with AR-V7 positive patients, PSA decline time (72.04±66.92 vs. 190.11±102.44, P=0.000), PSA non-response rate (6.12% vs. 21.4%, P=0.040) are significantly lower in AR-V7 negative patients. PSA PFS(489.17±269.39 vs. 130.56±120, P=0.010), Clinical PFS (551.91±322.05 vs. 261.44±200.85, P=0.018), and Imaging PFS (523.7±223.28 vs. 247.56±202.80, P=0.003) are significantly longer in AR-V7 negative patients. Conclusion Expression status of AR-V7 is related to the response of abiraterone treatment and the prognosis of the patients. Key words: Prostatic neoplasms; Castration resistant prostate cancer; Androgen receptor splice variant 7(AR-V7); Abiraterone

Leydig-like cells derived from reprogrammed human foreskin fibroblasts by CRISPR/dCas9 increase the level of serum testosterone in castrated male rats.

In the past few years, Leydig cell (LC) transplantation has been regarded as an effective strategy for providing physiological patterns of testosterone in vivo. Recently, we have successfully converted human foreskin fibroblasts (HFFs) into functional Leydig‐like cells (iLCs) in vitro by using the CRISPR/dCas9 system, which shows promising potential for seed cells. However, it is not known whether the reprogrammed iLCs can survive or restore serum testosterone levels in vivo. Therefore, in this study, we evaluate whether reprogrammed iLCs can restore the serum testosterone levels of castrated rats when they are transplanted into the fibrous capsule. We first developed the castrated Sprague Dawley rat model through bilateral orchiectomy and subsequently injected extracellular matrix gel containing transplanted cells into the fibrous capsule of castrated rats. Finally, we evaluated dynamic serum levels of testosterone and luteinizing hormone (LH) in castrated rats, the survival of implanted iLCs, and the expression levels of Leydig steroidogenic enzymes by immunofluorescence staining and Western blotting. Our results demonstrated that implanted iLCs could partially restore the serum testosterone level of castrated rats, weakly mimic the role of adult Leydig cells in the hypothalamic‐pituitary‐gonadal axis for a short period, and survive and secrete testosterone, through 6 weeks after transplantation. Therefore, this study may be valuable for treating male hypogonadism in the future.

Effect of cooling rate on microstructure and mechanical properties of CB2 tempered martensitic steel

The cooling rate of casting has a significant effect on microstructure and mechanical properties of castings. The 9Cr-1.5Mo-1Co cast steel, referred to as CB2, is one of the most promising alloys for various cast components to be used under ultra-supercritical conditions. In this study, HRTEM, SEM, and XRD methods were used for qualitative and quantitative analyses of contents, phases, and sizes of the inclusions and precipitates, as well as microstructure observation of the tempered martensitic steel at different cooling rates. Traditional tensile tests were conducted to characterize the material mechanical properties. Results show that the size of the boron nitride and precipitate, the width of the martensite lath and the content of the S-ferrite are reduced as the cooling rate increases from 5-8 °C-min-1 (CB2-S steel) to 50-60 °C-min-1 (CB2-F steel). The precipitates are mainly composed of M23C6 and a small amount of M3C. The average diameters of the M23C6 precipitates in CB2-F and CB2-S are 202 nm and 209 nm, respectively. The inclusions are mainly composed of BN, Al2O3 and MnO2, and the inclusion ratios are 0.1969% for the CB2-F and 0.06556% for CB2-S steel. The average martensite lath widths of CB2-F and CB2-S steels are 289 nm and 301 nm, respectively. Furthermore, the M3C having a diameter of about 150 nm and a thickness of 20 nm is observed in the 8-ferrite of the tempered CB2-S steel. The presence of the 8-ferrite reduces the precipitation strengthening and dislocation density in CB2-S steel. In addition, the lower cooling rate stabilizes the 8-ferrite structure at room temperature.

Extending Burk Dehority's Perspectives on the Role of Ciliate Protozoa in the Rumen.

Dr. Burk Dehority was an international expert on the classification and monoculture of ruminal ciliated protozoa. We have summarized many of the advancements in knowledge from his work but also in his scientific way of thinking about interactions of ruminal ciliates with the entire rumen microbial community and animal host. As a dedication to his legacy, an electronic library of high-resolution images and video footage catalogs numerous species and techniques involved in taxonomy, isolation, culture, and ecological assessment of ruminal ciliate species and communities. Considerable promise remains to adapt these landmark approaches to harness eukaryotic cell signaling technology with genomics and transcriptomics to assess cellular mechanisms regulating growth and responsiveness to ruminal environmental conditions. These technologies can be adapted to study how protozoa interact (both antagonism and mutualism) within the entire ruminal microbiota. Thus, advancements and limitations in approaches used are highlighted such that future research questions can be posed to study rumen protozoal contribution to ruminant nutrition and productivity.

Global chemical effects of the microbiome include new bile-acid conjugations.

A mosaic of cross-phylum chemical interactions occurs between all metazoans and their microbiomes. A number of molecular families that are known to be produced by the microbiome have a marked effect on the balance between health and disease1-9. Considering the diversity of the human microbiome (which numbers over 40,000operational taxonomic units10), the effect of the microbiome on the chemistry of an entire animal remains underexplored. Here we use mass spectrometry informatics and data visualization approaches11-13 to provide an assessment of the effects of the microbiome on the chemistry of an entire mammal by comparing metabolomics data from germ-free and specific-pathogen-free mice. We found that the microbiota affects the chemistry of all organs. This included the amino acid conjugations of host bile acids that were used to producephenylalanocholic acid, tyrosocholic acid and leucocholic acid, which have not previously been characterized despite extensive research on bile-acid chemistry14. These bile-acid conjugates were also found in humans, and were enriched in patients with inflammatory bowel disease or cystic fibrosis. These compounds agonized the farnesoidX receptor in vitro, and mice gavaged with the compounds showed reduced expression of bile-acid synthesis genes in vivo. Further studies are required to confirm whether these compounds have a physiological role in the host, and whether they contribute to gut diseases that are associated with microbiome dysbiosis.

High-quality, high-volume spay-neuter: Access to care and the challenge to private practitioners.

Low-cost spay-neuter clinics were first established nearly 50 years ago in response to the numbers of dogs and cats euthanized in animal shelters. Since then, high-quality, high-volume spay-neuter (HQHVSN) clinics have been established throughout the USA and have contributed to a significant reduction in animal euthanasia in shelters. These clinics, specializing in efficient systems and surgical techniques, provide an avenue for clients who cannot afford surgery to sterilize their pets. These clinics have also helped animal shelters with limited financial resources implement spay-neuter prior to adoption policies. The veterinary profession's reaction to HQHVSN clinics has been mixed; some practitioners question the quality of care provided or fear the loss of clientele, while others recognize the need for such clinics and support their development. As veterinary education has become more expensive and veterinary care more sophisticated, the cost of veterinary care, including spay-neuter surgery, has risen. With increasing costs, the numbers of pets that receive little or no veterinary care has increased. Indeed, a 2018 survey by the Access to Veterinary Care Coalition documented that more than 25% of pet-owning households in the USA experienced difficulties obtaining veterinary care for their pets and the most frequent barrier was financial. This review looks at the reality of HQHVSN clinics and what this means for the private practitioner. By adopting similar systems and techniques that lower the cost of spay-neuter surgery, practitioners could potentially pass on cost savings to clients. Moreover, the same principles may be applied to other aspects of basic care to further address access to care issues.

Large-scale cell-type-specific imaging of protein synthesis in a vertebrate brain.

Despite advances in methods to detect protein synthesis, it has not been possible to measure endogenous protein synthesis levels in vivo in an entire vertebrate brain. We developed a transgenic zebrafish line that allows for cell-type-specific labeling and imaging of nascent proteins in the entire animal. By replacing leucine with glycine in the zebrafish MetRS-binding pocket (MetRS-L270G), we enabled the cell-type-specific incorporation of the azide-bearing non-canonical-amino-acid azidonorleucine (ANL) during protein synthesis. Newly synthesized proteins were then labeled via 'click chemistry'. Using a Gal4-UAS-ELAV3 line to express MetRS-L270G in neurons, we measured protein synthesis intensities across the entire nervous system. We visualized endogenous protein synthesis and demonstrated that seizure-induced neural activity results in enhanced translation levels in neurons. This method allows for robust analysis of endogenous protein synthesis in a cell-type-specific manner, in vivo at single-cell resolution.