Extensive dynamic changes in the human transcriptome and its circadian organization during prolonged bed rest.

Physiological and molecular processes including the transcriptome change across the 24-h day, driven by molecular circadian clocks and behavioral and systemic factors. It is not known how the temporal organization of the human transcriptome responds to a long-lasting challenge. This may, however, provide insights into adaptation, disease, and recovery. We investigated the human 24-h time series transcriptome in 20 individuals during a 90-day constant bed rest protocol. We show that the protocol affected 91% of the transcriptome with 76% of the transcriptome still affected after 10 days of recovery. Dimensionality-reduction approaches revealed that many affected transcripts were associated with mRNA translation and immune function. The number, amplitude, and phase of rhythmic transcripts, including clock genes, varied significantly across the challenge. These findings of long-lasting changes in the temporal organization of the transcriptome have implications for understanding the mechanisms underlying health consequences of conditions such as microgravity and bed rest.

HIV-2 mediated effects on target and bystander cells induce plasma proteome remodeling.

Despite low or undetectable plasma viral load, people living with HIV-2 (PLWH2) typically progress toward AIDS. The driving forces behind HIV-2 disease progression and the role of viremia are still not known, but low-level replication in tissues is believed to play a role. To investigate the impact of viremic and aviremic HIV-2 infection on target and bystander cell pathology, we used data-independent acquisition mass spectrometry to determine plasma signatures of tissue and cell type engagement. Proteins derived from target and bystander cells in multiple tissues, such as the gastrointestinal tract and brain, were detected at elevated levels in plasma of PLWH2, compared with HIV negative controls. Moreover, viremic HIV-2 infection appeared to induce enhanced release of proteins from a broader range of tissues compared to aviremic HIV-2 infection. This study expands the knowledge on the link between plasma proteome remodeling and the pathological cell engagement in tissues during HIV-2 infection.

Soluble ACE2 correlates with severe COVID-19 and can impair antibody responses.

Identifying immune modulators that impact neutralizing antibody responses against severe acute respiratory syndrome coronavirus type 2 (SARS-CoV-2) is of great relevance. We postulated that high serum concentrations of soluble angiotensin-converting enzyme 2 (sACE2) might mask the spike and interfere with antibody maturation toward the SARS-CoV-2-receptor-binding motif (RBM). We tested 717 longitudinal samples from 295 COVID-19 patients and showed a 2- to 10-fold increase of enzymatically active sACE2 (a-sACE2), with up to 1 µg/mL total sACE2 in moderate and severe patients. Fifty percent of COVID-19 sera inhibited ACE2 activity, in contrast to 1.3% of healthy donors and 4% of non-COVID-19 pneumonia patients. A mild inverse correlation of a-sACE2 with RBM-directed serum antibodies was observed. In silico, we show that sACE2 concentrations measured in COVID-19 sera can disrupt germinal center formation and inhibit timely production of high-affinity antibodies. We suggest that sACE2 is a biomarker for COVID-19 and that soluble receptors may contribute to immune suppression informing vaccine design.

Biomonitoring of glyphosate and aminomethylphosphonic acid: Current insights and future perspectives.

Glyphosate is one of the most widely used herbicides globally, raising concerns about its potential impact on human health. Biomonitoring studies play a crucial role in assessing human exposure to glyphosate and providing valuable insights into its distribution and metabolism in the body. This review aims to summarize the current trends and future perspectives in biomonitoring of glyphosate and its major degradation product of aminomethylphosphonic acid (AMPA). A comprehensive literature search was conducted, focusing on studies published between January 2000 and December 2022. The findings demonstrated that glyphosate and AMPA have been reported in different human specimens with urine as the dominance. Sample pretreatment techniques of solid-phase and liquid-liquid extractions coupled with liquid/gas chromatography-tandem mass spectrometry have achieved matrix elimination and accurate analysis. We also examined and compared the exposure characteristics of these compounds among different regions and various populations, with significantly higher levels of glyphosate and AMPA observed in Asian populations and among occupational groups. The median urinary concentration of glyphosate in children was 0.54 ng/mL, which was relatively higher than those in women (0.28 ng/mL) and adults (0.12 ng/mL). It is worth noting that children may exhibit increased susceptibility to glyphosate exposure or have different exposure patterns compared to women and adults. A number of important perspectives were proposed in order to further facilitate the understanding of health effects of glyphosate and AMPA, which include, but are not limited to, method standardization, combined exposure assessment, attention for vulnerable populations, long-term exposure effects and risk communication and public awareness.

Challenges associated with vascular access through the common carotid artery during embalming procedures in humans: A technical note.

BACKGROUND: The teaching of anatomy is a key component in the training of physicians, and the foundation of this teaching is the human body, which must be properly prepared to be used as a teaching aid. Due to a lack of modern literature on this topic, we decided to write a technical note discussing access to the carotid artery. MATERIALS AND METHODS: We pre-qualified 43 donor bodies for the study. The bodies had to meet standards such as no signs of post-mortem decomposition, preservation of body integrity, and the absence of known infections. Carotid artery access was performed based on descriptions of the types of vascular access performed in surgery and our own observations. RESULTS: We consider carotid artery access to be a convenient option due to its ease of location. When performed correctly and with attention to the surrounding structures, it is relatively low in tissue trauma, which translates into a higher quality of preparation. Data analysis has revealed several factors that can have a significant impact on the success of the embalming procedure. CONCLUSIONS: Proper execution of minimally invasive access to the common carotid artery minimizes tissue damage and ensures a high success rate of the procedure. Knowledge of the types of vascular access is essential for preparing the highest quality specimens.

In silico analysis and theratyping of an ultra-rare CFTR genotype (W57G/A234D) in primary human rectal and nasal epithelial cells.

Mutation targeted therapy in cystic fibrosis (CF) is still not eligible for all CF subjects, especially for cases carrying rare variants such as the CFTR genotype W57G/A234D (c.169T>G/c.701C>A). We performed in silico analysis of the effects of these variants on protein stability, which we functionally characterized using colonoids and reprogrammed nasal epithelial cells. The effect of mutations on cystic fibrosis transmembrane conductance regulator (CFTR) protein was analyzed by western blotting, forskolin-induced swelling (FIS), and Ussing chamber analysis. We detected a residual CFTR function that increases following treatment with the CFTR modulators VX661±VX445±VX770, correlates among models, and is associated with increased CFTR protein levels following treatment with CFTR correctors. In vivo treatment with VX770 reduced sweat chloride concentration to non-CF levels, increased the number of CFTR-dependent sweat droplets, and induced a 6% absolute increase in predicted FEV1% after 27 weeks of treatment indicating the relevance of theratyping with patient-derived cells in CF.

Neonicotinoid insecticides and their metabolites: Specimens tested, analytical methods and exposure characteristics in humans.

The use of neonicotinoid insecticides (NEOs) has been rising globally due to their broad-spectrum insecticidal activity, unique mode of neurotoxic action and presumed low mammalian toxicity. Given their growing ubiquity in the environment and neurological toxicity to non-target mammals, human exposure to NEOs is flourishing and now becomes a big issue. In the present work, we demonstrated that 20 NEOs and their metabolites have been reported in different human specimens with urine, blood and hair as the dominance. Sample pretreatment techniques of solid-phase and liquid-liquid extractions coupled with high performance liquid chromatography-tandem mass spectrometry have successfully achieved matrix elimination and accurate analysis. We also discussed and compared exposure characteristics of these compounds among types of specimens and different regions. A number of important knowledge gaps were also identified in order to further facilitate the understanding of health effects of NEO insecticides, which include, but are not limited to, identification and use of neuro-related human biological samples for better elucidating neurotoxic action of NEO insecticides, adoption of advanced non-target screening analysis for a whole picture in human exposure, and expanding investigations to cover non-explored but NEO-used regions and vulnerable populations.

Single-cell transcriptomes underscore genetically distinct tumor characteristics and microenvironment for hereditary kidney cancers.

Our understanding of how each hereditary kidney cancer adapts to its tissue microenvironment is incomplete. Here, we present single-cell transcriptomes of 108,342 cells from patient specimens including from six hereditary kidney cancers. The transcriptomes displayed distinct characteristics of the cell of origin and unique tissue microenvironment for each hereditary kidney cancer. Of note, hereditary leiomyomatosis and renal cell carcinoma (HLRCC)-associated kidney cancer retained some characteristics of proximal tubules, which were completely lost in lymph node metastases and present as an avascular tumor with suppressed T cells and TREM2-high macrophages, leading to immune tolerance. Birt-Hogg-Dubé (BHD)-associated kidney cancer exhibited transcriptomic intratumor heterogeneity (tITH) with increased characteristics of intercalated cells of the collecting duct and upregulation of FOXI1-driven genes, a critical transcription factor for collecting duct differentiation. These findings facilitate our understanding of how hereditary kidney cancers adapt to their tissue microenvironment.

Treatment of acromegaly by rosiglitazone via upregulating 15-PGDH in both pituitary adenoma and liver.

Rosiglitazone, a synthetic peroxisome proliferator-activated receptor γ (PPARγ) ligand, has been reported to reduce growth hormone (GH) and insulin-like growth factor-1 (IGF-1) in 10 patients with acromegaly. However, the mechanisms remain unknown. Here, we reveal that PPARγ directly enhances 15-hydroxyprostaglandin dehydrogenase (15-PGDH) expression, whose expression is decreased and negatively correlates with tumor size in acromegaly. Rosiglitazone decreases GH production and promotes apoptosis and autophagy in GH3 and primary somatotroph adenoma cells and suppresses hepatic GH receptor (GHR) expression and IGF-1 secretion in HepG2 cells. Activating the PGE2/cAMP/PKA pathway directly increases GHR expression. Rosiglitazone suppresses tumor growth and decreases GH and IGF-1 levels in mice inoculated subcutaneously with GH3 cells. The above effects are all dependent on 15-PGDH expression. Rosiglitazone as monotherapy effectively decreases GH and IGF-1 levels in all nineteen patients with active acromegaly. Evidence suggests that rosiglitazone may be an alternative pharmacological approach for acromegaly by targeting both pituitary adenomas and liver.

Characterization of SARS-CoV-2 nucleocapsid protein reveals multiple functional consequences of the C-terminal domain.

Nucleocapsid (N) encoded by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) plays key roles in the replication cycle and is a critical serological marker. Here, we characterize essential biochemical properties of N and describe the utility of these insights in serological studies. We define N domains important for oligomerization and RNA binding and show that N oligomerization provides a high-affinity RNA-binding platform. We also map the RNA-binding interface, showing protection in the N-terminal domain and linker region. In addition, phosphorylation causes reduction of RNA binding and redistribution of N from liquid droplets to loose coils, showing how N-RNA accessibility and assembly may be regulated by phosphorylation. Finally, we find that the C-terminal domain of N is the most immunogenic, based on antibody binding to patient samples. Together, we provide a biochemical description of SARS-CoV-2 N and highlight the value of using N domains as highly specific and sensitive diagnostic markers.

Respond of the different human cranial bones to pin-type head fixation device.

BACKGROUND: At this juncture, there is no consensus in the literature for the use and the safety of pin-type head holders in cranial procedures. METHODS: The present analysis of the bone response to the fixation of the instrument provides data to understand its impact on the entire skull as well as associated complications. An experimental study was conducted on fresh-frozen human specimens to analyze the puncture hole due to the fixation of each single pin of the pin-type head holder. Cone-beam CT images were acquired to measure the diameter of the puncture hole caused by the instrument according to several parameters: the pin angle, the clamping force, and different neurosurgical approaches most clinically used. RESULTS: The deepest hole, 2.67 ± 0.27 mm, was recorded for a 35° angle and a clamping force of 270 N at the middle fossa approach. The shallowest hole was 0.62 ± 0.22 mm for the 43° angle with a pinning force of 180 N in the pterional approach. The pterional approach had a significantly different effect on the depth of the puncture hole compared with the middle fossa craniotomy for 270 N pinning at 35° angle. The puncture hole measured with the 43° angle and 180 N force in prone position is significantly different from the other approaches with the same force. CONCLUSIONS: These results could lead to recommendations about the use of the head holder depending on the patient's history and cranial thickness to reduce complications associated with the pin-type head holder during clinical applications.

Lag-Screw Osteosynthesis in Thoracolumbar Pincer Fractures.

STUDY DESIGN: Biomechanical. OBJECTIVE: This study evaluates the biomechanical properties of lag-screws used in vertebral pincer fractures at the thoracolumbar junction. METHODS: Pincer fractures were created in 18 bisegmental human specimens. The specimens were assigned to three groups depending on their treatment perspective, either bolted, with the thread positioned in the cortical or cancellous bone, or control. The specimens were mounted in a servo-hydraulic testing machine and loaded with a 500 N follower load. They were consecutively tested in 3 different conditions: intact, fractured, and bolted/control. For each condition 10 cycles in extension/flexion, torsion, and lateral bending were applied. After each tested condition, a computed tomography (CT) scan was performed. Finally, an extension/flexion fatigue loading was applied to all specimens. RESULTS: Biomechanical results revealed a nonsignificant increase in stiffness in extension/flexion of the fractured specimens compared with the intact ones. For lateral bending and torsion, the stiffness was significantly lower. Compared with the fractured specimens, no changes in stiffness due to bolting were discovered. CT scans showed an increasing fracture gap during axial loading both in extension/flexion, torsion, and lateral bending in the control specimens. In bolted specimens, the anterior fragment was approximated, and the fracture gap nullified. This refers to both the cortical and the cancellous thread positions. CONCLUSION: The results of this study concerning the effect of lag-screws on pincer fractures appear promising. Though there was little effect on stiffness, CT scans reveal a bony contact in the bolted specimens, which is a requirement for bony healing.

Effect of a formalin-based fixation method on bone mineral content in human ex-vivo specimens.

OBJECTIVE: Histopathology of formalin-fixated human ex-vivo specimens may be used as reference standard for evaluation of diagnostic index tests like CBCT or MRI. The aim was to estimate changes in bone mineral content (BMC) over time in human ex-vivo bone specimens fixated in a formalin-based solution for 24 h followed by storage in an alcohol-based medium for six months, assessed by dual-energy X-ray absorptiometry (DXA). METHODOLOGY: Bone specimens (n = 19) from human ex-vivo mandibles donated for science were included. BMC was measured by DXA before fixation (D0), after 24 h of immersion fixation in a formalin-based solution (D1), and hereafter every 30 days (M1-M6) during storage in a 30% ethanol-based storage medium for 6 months. Changes in BMC from D0 to D1 and from D0 to M6 were calculated and mean change in BMC estimated. RESULTS: Mean change in BMC from D0 to D1 was -0.73% (95% CI -1.75%; 0.29%), and from D0 to M6 -1.19% (95% CI -2.14%; -0.23%). CONCLUSIONS: No changes in BMC of ex-vivo human bone specimens were found after 24 h formalin-based immersion fixation. After six months storage in an ethanol-based medium, BMC mean loss of 1% was detected. In this range, changes in BMC are not clinically relevant.

Characterization of tissue properties in epidural needle insertion on human specimen and synthetic materials.

The force experienced while inserting an 18-gauge Tuohy needle into the epidural space or dura is one of only two feedback components perceived by an anaesthesiologist to deduce the needle tip position in a patient's spine. To the best of the authors knowledge, no x-ray validated measurements of these forces are currently available to the public. A needle insertion force recording during an automated insertion of an 18-gauge Tuohy needle into human vertebral segments of four female donors was conducted. During the measurements, x-ray images were recorded simultaneously. The force peaks due to the penetration of the ligamentum supraspinale and ligamentum flavum were measured and compared to the measurements of an artificial patient phantom for a hybrid patient simulator. Based on these force peaks and the slope of the ligamentum interspinale, a mathematical model was developed. The model parameters were used to compare human specimens and artificial patient phantom haptics. The force peaks for the ligamenta supraspinale and flavum were 7.55 ± 3.63 N and 15.18 ± 5.71 N, respectively. No significant differences were found between the patient phantom and the human specimens for the force peaks and four of six physical model parameters. The patient phantom mimics the same resistive force against the insertion of an 18-gauge Tuohy needle. However, there was a highly significant (p < 0.001, effsize = 0.949 and p < 0.001, effsize = 0.896) statistical difference observed in the insertion depth where the force peaks of the ligamenta supraspinale and flavum were detected between the measurements on the human specimens and the patient phantom. Within this work, biomechanical evidence was identified for the needle insertion force into human specimens. The comparison of the measured values of the human vertebral segments and the artificial patient phantom showed promising results.

Obstruction of Small Arterioles in Patients with Critical Limb Ischemia due to Partial Endothelial-to-Mesenchymal Transition.

Critical limb ischemia (CLI) is a hazardous manifestation of atherosclerosis and treatment failure is common. Abnormalities in the arterioles might underlie this failure but the cellular pathobiology of microvessels in CLI is poorly understood. We analyzed 349 intramuscular arterioles in lower limb specimens from individuals with and without CLI. Arteriolar densities were 1.8-fold higher in CLI muscles. However, 33% of small (<20 µm) arterioles were stenotic and 9% were completely occluded. The lumens were closed by bulky, re-oriented endothelial cells expressing abundant N-cadherin that uniquely localized between adjacent and opposing endothelial cells. S100A4 and SNAIL1 were also expressed, supporting an endothelial-to-mesenchymal transition. SMAD2/3 was activated in occlusive endothelial cells and TGFß1 was increased in the adjacent mural cells. These findings identify a microvascular closure process based on mesenchymal transitions in a hyper-TGFß environment that may, in part, explain the limited success of peripheral artery revascularization procedures.

Rupture limit evaluation of human cerebral aneurysms wall: Experimental study.

BACKGROUND AND PURPOSE: Rupture risk of intracranial aneurysms is a major issue for public healthcare. A way to obtain an individual rupture risk assessment is a main objective of many research teams in the world. For many years, we have investigated the relationship between the mechanical properties of aneurysm wall tissues and the rupture risk. In this work, we try to go further and investigate rupture limit values. METHODS: Following surgical clipping, a specific conservation protocol was applied to aneurysmal tissues in order to preserve their mechanical properties. Thirty-nine intracranial aneurysms (27 females, 12 males) were tested using a uniaxial tensile test machine under physiological conditions, temperature, and saline isotonic solution. These represented 24 unruptured and 15 ruptured aneurysms. Stress/strain curves were then obtained for each sample, and a fitting algorithm was applied following a Yeoh hyperelastic model with 2 parameters. Moreover, uniaxial tensile tests were conducted until rupture of samples to obtain values of stress and strain rupture limit. RESULTS: The significant parameter a C2 of the hyperelastic Yeoh model, allowed us to classify samples' rigidity following the terminology we adopted in previous papers (Costalat et al., 2011; Sanchez et al., 2013): Soft, Stiff and Intermediate. Moreover, strain/stress rupture limit values were gathered and analyzed thanks to the tissue rigidity, the status of the aneurysm (initially ruptured or unruptured) and the gender of the patient. CONCLUSION: Strain rupture limit was found quite stable around 20% and seems not to be correlated with the status of the aneurysm (initially ruptured or unruptured), neither with the gender of the patient. However, stretch and stress rupture limit seems not to be independent on the rigidity. The study confirms that ruptured aneurysms mainly present a soft tissue and unruptured aneurysms present a stiff material.

Use of human specimens in research: the evolving United States regulatory, policy, and scientific landscape.

The use of human specimens in research has contributed to significant scientific and medical advancements. However, the development of sophisticated whole genome and informatics technologies and the increase in specimen and data sharing have raised new questions about the identifiability of specimens and the protection of participants in human specimen research. In the US, new regulations and policies are being considered to address these changes. This review discusses the current and proposed regulations as they apply to specimen research, as well as relevant policy discussions. It summarizes the ways that researchers and other stakeholders can provide their input to these discussions and policy development efforts. Input from all the stakeholders in specimen research will be essential for the development of policies that facilitate such research while at the same time protecting the rights and welfare of research participants.