Mesenchymal stem cell: keystone of the hematopoietic stem cell niche and a stepping-stone for regenerative medicine.

Mesenchymal stem cells (MSCs) are self-renewing precursor cells that can differentiate into bone, fat, cartilage, and stromal cells of the bone marrow. Recent studies suggest that MSCs themselves are critical for forming a niche that maintains hematopoietic stem cells (HSCs). The ease by which human MSC-like and stromal progenitor cells can be isolated from the bone marrow and other tissues has led to the rapid development of clinical investigations exploring their anti-inflammatory properties, tissue preservation capabilities, and regenerative potential. However, the identity of genuine MSCs and their specific contributions to these various beneficial effects have remained enigmatic. In this article, we examine the definition of MSCs and discuss the importance of rigorously characterizing their stem cell activity. We review their role and that of other putative niche constituents in the regulation of bone marrow HSCs. Additionally, how MSCs and their stromal progeny alter immune function is discussed, as well as potential therapeutic implications.

Resilient anatomy and local plasticity of naive and stress haematopoiesis.

The bone marrow adjusts blood cell production to meet physiological demands in response to insults. The spatial organization of normal and stress responses are unknown owing to the lack of methods to visualize most steps of blood production. Here we develop strategies to image multipotent haematopoiesis, erythropoiesis and lymphopoiesis in mice. We combine these with imaging of myelopoiesis1 to define the anatomy of normal and stress haematopoiesis. In the steady state, across the skeleton, single stem cells and multipotent progenitors distribute through the marrow enriched near megakaryocytes. Lineage-committed progenitors are recruited to blood vessels, where they contribute to lineage-specific microanatomical structures composed of progenitors and immature cells, which function as the production sites for each major blood lineage. This overall anatomy is resilient to insults, as it was maintained after haemorrhage, systemic bacterial infection and granulocyte colony-stimulating factor (G-CSF) treatment, and during ageing. Production sites enable haematopoietic plasticity as they differentially and selectively modulate their numbers and output in response to insults. We found that stress responses are variable across the skeleton: the tibia and the sternum respond in opposite ways to G-CSF, and the skull does not increase erythropoiesis after haemorrhage. Our studies enable in situ analyses of haematopoiesis, define the anatomy of normal and stress responses, identify discrete microanatomical production sites that confer plasticity to haematopoiesis, and uncover unprecedented heterogeneity of stress responses across the skeleton.

In situ mapping identifies distinct vascular niches for myelopoiesis.

In contrast to nearly all other tissues, the anatomy of cell differentiation in the bone marrow remains unknown. This is owing to a lack of strategies for examining myelopoiesis-the differentiation of myeloid progenitors into a large variety of innate immune cells-in situ in the bone marrow. Such strategies are required to understand differentiation and lineage-commitment decisions, and to define how spatial organizing cues inform tissue function. Here we develop approaches for imaging myelopoiesis in mice, and generate atlases showing the differentiation of granulocytes, monocytes and dendritic cells. The generation of granulocytes and dendritic cells-monocytes localizes to different blood-vessel structures known as sinusoids, and displays lineage-specific spatial and clonal architectures. Acute systemic infection with Listeria monocytogenes induces lineage-specific progenitor clusters to undergo increased self-renewal of progenitors, but the different lineages remain spatially separated. Monocyte-dendritic cell progenitors (MDPs) map with nonclassical monocytes and conventional dendritic cells; these localize to a subset of blood vessels expressing a major regulator of myelopoiesis, colony-stimulating factor 1 (CSF1, also known as M-CSF)1. Specific deletion of Csf1 in endothelium disrupts the architecture around MDPs and their localization to sinusoids. Subsequently, there are fewer MDPs and their ability to differentiate is reduced, leading to a loss of nonclassical monocytes and dendritic cells during both homeostasis and infection. These data indicate that local cues produced by distinct blood vessels are responsible for the spatial organization of definitive blood cell differentiation.

Comprehensive analysis of gastrointestinal side effects in COVID-19 patients undergoing combined pharmacological treatment with azithromycin and hydroxychloroquine: a systematic review and network meta-analysis.

During the COVID-19 pandemic, several drugs were repositioned and combined to quickly find a way to mitigate the effects of the infection. However, the adverse effects of these combinations on the gastrointestinal tract are unknown. We aimed investigate whether Hydroxychloroquine (HD), Azithromycin (AZ), and Ivermectin (IV) used in combination for the treatment of COVID-19, can lead to the development of gastrointestinal disorders. This is a systematic review and network meta-analysis conducted using Stata and Revman software, respectively. The protocol was registered with PROSPERO (CRD42023372802). A search of clinical trials in Cochrane Library databases, Embase, Web of Science, Lilacs, PubMed, Scopus and Clinicaltrials.gov conducted on November 26, 2023. The eligibility of the studies was assessed based on PICO criteria, including trials that compared different treatments and control group. The analysis of the quality of the evidence was carried out according to the GRADE. Six trials involving 1,686 COVID-19 patients were included. No trials on the association of HD or AZ with IV met the inclusion criteria, only studies on the association between HD and AZ were included. Nausea, vomiting, diarrhea, abdominal pain and increased transaminases were related. The symptoms of vomiting and nausea were evaluated through a network meta-analysis, while the symptom of abdominal pain was evaluated through a meta-analysis. No significant associations with these symptoms were observed for HD, AZ, or their combination, compared to control. Low heterogeneity and absence of inconsistency in indirect and direct comparisons were noted. Limitations included small sample sizes, varied drug dosages, and potential publication bias during the pandemic peak. This review unveils that there are no associations between gastrointestinal adverse effects and the combined treatment of HD with AZ in the management of COVID-19, as compared to either the use of a control group or the administration of the drugs individually, on the other hand, highlighting the very low or low certainty of evidence for the evaluated outcomes. To accurately conclude the absence of side effects, further high-quality randomized studies are needed.

Molecular detection of Hepatozoon species (Apicomplexa: Hepatozoidae) infecting snakes in the Northeastern region of Argentina.

The genus Hepatozoon Miller (1908) contains a wide range of obligate parasitic organisms with complex life cycles involving vertebrates and hematophagous invertebrates. Despite over 300 species being described, only a small percentage has been characterized in snakes using morphological and molecular techniques. The prevalence of these parasites in snakes is significant, highlighting the need for molecular descriptions in such elusive hosts. Thus, the objective of this study was to determine molecularly the presence of Hepatozoon species in snakes from the Northeastern region of Argentina. Thirty-two specimens of eight snake species (Bothrops alternatus, Dryophylax hypoconia, Erythrolamprus jaegeri coralliventris, Erythrolamprus poecilogyrus, Erythrolamprus semiaureus, Philodryas olfersii latirostris, Pseudablabes (ex Philodryas) patagoniensis and Palusophis (ex Mastigodryas) bifossatus were collected and examined. PCR analysis of the 18S rRNA locus detected four samples (12% prevalence) positive for the presence of Hepatozoon DNA. Phylogenetic analysis positioned the 18S rRNA Hepatozoon sequences obtained in three different clades, one with Hepatozoon musa, another with sequences of Hepatozoon cuestensis, while the third was placed as a sister taxon to a clade including Hepatozoon cevapii and Hepatozoon massardi. This study presents the first documentation of Hepatozoon infecting snakes in Argentina, thereby expanding their distribution within southern South America. Additionally, B. alternatus and Pa. bifossatus are reported as new hosts of Hepatozoon.

Open Bankart repair plus inferior capsular shift versus isolated arthroscopic Bankart repair in collision athletes with recurrent anterior shoulder instability: a prospective study.

BACKGROUND: Open Bankart repair plus inferior capsular shift and isolated arthroscopic Bankart repair have never been prospectively compared under the concept of glenoid track in collision athletes with recurrent anterior shoulder instability. The aim of this study was to compare the functional outcomes, range of motion, and recurrence rate between these 2 surgical techniques. We hypothesized that open Bankart repair plus inferior capsular shift would provide similar functional outcomes to isolated arthroscopic Bankart repair but with a lower recurrence rate. METHODS: A prospective cohort study was conducted with 86 collision athletes divided into 2 groups of 43 patients each. All patients had a subcritical glenoid bone loss ≤13.5% and an on-track Hill Sachs lesion. The average follow-up was 66 (60-93) months for the open group and 68 (60-97) months for the arthroscopic group. The primary functional outcomes of each group were evaluated at baseline, 6 months, 1 year and for a minimum of 5 years after surgery. The functional outcomes were also compared between the 2 groups. The assessment tools included the Western Ontario Shoulder Instability Index (WOSI) score and American Shoulder and Elbow Surgeons scale (ASES) score. In addition, recurrent instability and range of motion were also evaluated. RESULTS: In each group, there were significant differences in Western Ontario Shoulder Instability Index score and American Shoulder and Elbow Surgeons scale score between the pre and postoperative periods. There were no differences between the groups at the end of follow-up (P = .47 and .22). Three dislocations (6.9%) in the open group and 10 dislocations (23.2%) in the arthroscopic group were reported showing significant differences (P = .012). In addition, there were no differences in range of motion between pre and postoperative periods for each group as well as between them. CONCLUSION: We found no differences in functional outcomes and range of motion between the 2 groups. The recurrence rate was significantly higher in the arthroscopic group. We recommend performing open Bankart repair plus inferior capsular shift as a treatment alternative in collision athletes with recurrent anterior shoulder instability.

Similar outcomes in collision athletes with subcritical glenoid bone loss and on-Track Hill Sachs lesion versus off-track Hill Sachs lesion managed with open Bankart repair plus inferior capsular shift.

INTRODUCTION: Open Bankart repair plus inferior capsular shift has not yet been tested under the concept of glenoid track as a predictor of failure. The aim of this study was to compare the subjective and objective outcomes in collision athletes with subcritical glenoid bone loss and on-track Hill Sachs lesions versus those with off-track Hill Sachs lesions, all treated with open Bankart repair. METHODS: Two study groups were created: 50 patients had on-track Hill Sachs lesions, while 38 had off-track lesions. The subcritical glenoid bone loss was ≤ 10%. A minimum follow-up period of 3 years was established. Preoperative and postoperative evaluation of each group and between them was performed. The Western Ontario Shoulder Instability Index score and the American Shoulder and Elbow Surgeons scale were used to assess subjective outcomes. Recurrence rate, range of motion and return to sport were evaluated as objective outcomes. RESULTS: Significant differences were reported in the WOSI and ASES scores between preoperative and postoperative values in each group. There were no significant differences between the two groups (p-value = 0.36 and 0.71). Three dislocations (6%) in the on-track group and 3 (7.8%) in the off-track group were recorded, showing no differences between the two groups (p-value = 0.83). There were no differences in ROM between pre- and post-operatively in each group or when comparing the two groups. CONCLUSIONS: We found no differences between the outcomes of the two groups. According to the surgeon's preference, we recommend performing open Bankart repair plus inferior capsular shift as a treatment alternative in collision athletes with SGBL ≤ 10% independently of the type of Hill Sachs lesion.

Comparison between arthroscopic Bankart repair versus arthroscopic Bankart/SLAP lesion repair in limited-contact athletes with type V SLAP lesion. A prospective cohort study.

PURPOSE: The aim of this study was to compare the functional outcomes, recurrence rate, range of motion (ROM) and return to sports activities between arthroscopic Bankart repair (ABR) versus arthroscopic Bankart/SLAP repair (ABR/S) in limited contact-athletes with a type V SLAP lesion in the scenario of recurrent anterior shoulder instability (RASI). Our hypothesis was that there is no difference between the two treatments. METHODS: Two groups of 45 limited-contact athletes with type V SLAP lesion were created. Group 1 underwent an arthroscopic Bankart repair, while group 2 had an arthroscopic Bankart/SLAP repair. The minimum follow-up period was 2 years. The WOSI and ASES scores were used to assess primary functional outcomes. Recurrence rate, ROM and return to sport were also evaluated. RESULTS: Significant differences were reported in the WOSI and ASES scores pre- and post-operatively in each group. There were no significant differences between the two groups (P = 0.78 and 0.43). We reported 4 recurrences (8.8 %) in group 1 and 5 (11.1 %) in group 2, with no difference between them (P = 0.62). There were no significant differences between the range of motion of each of the groups as well as between them. More than 90% of the athletes in both groups returned to their previous sporting activities. CONCLUSIONS: Limited-contact athletes with RASI who have a type V SLAP lesion as their primary diagnosis can be treated using either ABR or ABR/S with equal efficacy. Both treatment alternatives preserve athlete's function, stability, ROM and return to sport.

A new hyperelastic lookup table for RT-DC.

Real-time deformability cytometry (RT-DC) is an established method that quantifies features like size, shape, and stiffness for whole cell populations on a single-cell level in real-time. A lookup table (LUT) disentangles the experimentally derived steady-state cell deformation and the projected area to extract the cell stiffness in the form of the Young's modulus. So far, two lookup tables exist but are limited to simple linear material models and cylindrical channel geometries. Here, we present two new lookup tables for RT-DC based on a neo-Hookean hyperelastic material numerically derived by simulations based on the finite element method in square and cylindrical channel geometries. At the same time, we quantify the influence of the shear-thinning behavior of the surrounding medium on the stationary deformation of cells in RT-DC and discuss the applicability and impact of the proposed LUTs regarding past and future RT-DC data analysis. Additionally, we provide insights about the cell strain and stresses, as well as the influence resulting from the rotational symmetric assumption on the cell deformation and volume estimation. The new lookup tables and the numerical cell shapes are made freely available.

Experimental and Theoretical Investigation of the Reaction of NH2 with NO at Very Low Temperatures.

The first experimental study of the low-temperature kinetics of the gas-phase reaction between NH2 and NO has been performed. A pulsed laser photolysis-laser-induced fluorescence technique was used to create and monitor the temporal decay of NH2 in the presence of NO. Measurements were carried out over the temperature range of 24-106 K, with the low temperatures achieved using a pulsed Laval nozzle expansion. The negative temperature dependence of the reaction rate coefficient observed at higher temperatures in the literature continues at these lower temperatures, with the rate coefficient reaching 3.5 × 10-10 cm3 molecule-1 s-1 at T = 26 K. Ab initio calculations of the potential energy surface were combined with rate theory calculations using the MESMER software package in order to calculate and predict rate coefficients and branching ratios over a wide range of temperatures, which are largely consistent with experimentally determined literature values. These theoretical calculations indicate that at the low temperatures investigated for this reaction, only one product channel producing N2 + H2O is important. The rate coefficients determined in this study were used in a gas-phase astrochemical model. Models were run over a range of physical conditions appropriate for cold to warm molecular clouds (10 to 30 K; 104 to 106 cm-3), resulting in only minor changes (<1%) to the abundances of NH2 and NO at steady state. Hence, despite the observed increase in the rate at low temperatures, this mechanism is not a dominant loss mechanism for either NH2 or NO under dark cloud conditions.

Ab Initio and Statistical Rate Theory Exploration of the CH (X2Π) + OCS Gas-Phase Reaction.

The first theoretical results regarding the gas-phase reaction mechanism and kinetics of the CH (X2Π) + OCS reaction are presented here. This reaction has a proposed importance in the removal of OCS in regions of the interstellar medium (ISM) and has the potential to form the recently observed HCS/HSC isomers, with both constitutional isomers having recently been observed in the L483 molecular cloud in a 40:1 ratio. Statistical rate theory simulations were performed on stationary points along the reaction potential energy surface (PES) obtained from ab initio calculations at the RO-CCSD(T)/aug-cc-pV(Q+d)Z//M06-2X-D3/aug-cc-pV(Q+d)Z level of theory over the temperature and total density range of 150-3000 K and 1011-1024 cm-3, respectively, using a Master Equation analysis. Exploration of the reaction potential energy surface revealed that all three pathways identified to create CS + HCO products required surmounting barriers of 16.5 kJ mol-1 or larger when CH approached the oxygen side of OCS, rendering this product formation negligible below 1000 K, and certainly under low-temperature ISM conditions. In contrast, when CH approaches the sulfur side of OCS, only submerged barriers are found along the reaction potential energy surface to create HCCO + S or CO + HCS, both of which are formed via a strongly bound OCC(H)S intermediate (-358.9 kJ mol-1). Conversion from HCS to HSC is possible via a barrier of 77.8 kJ mol-1, which is still -34.1 kJ mol-1 below the CH + OCS entrance channel. No direct route from CH + OCS to H + CO + CS was found from our ab initio calculations. Rate theory simulations suggest that the reaction has a strong negative temperature dependence, in accordance with the barrierless addition of CH to the sulfur side of OCS. Product branching fractions were also determined from MESMER simulations over the same temperature and total density range. The product branching fraction of CO + HCS reduces from 79% at 150 K to 0.0% at 800 K, while that of HCS dissociation to H + CS + CO increases from 22% at 150 K to 100% at 800 K. The finding of CO + HCS as the major product at the low temperatures relevant to the ISM, instead of H + CS + CO, is in opposition to the current supposition used in the KIDA database and should be adapted in astrochemical models as another source of the HCS isomer.

Ecological factors shaping the ectoparasite community assembly of the Azara's Grass Mouse, Akodon azarae (Rodentia: Cricetidae).

Parasites are integral members of the global biodiversity. They are useful indicators of environmental stress, food web structure and diversity. Ectoparasites have the potential to transmit vector-borne diseases of public health and veterinary importance and to play an important role in the regulation and evolution of host populations. The interlinkages between hosts, parasites and the environment are complex and challenging to study, leading to controversial results. Most previous studies have been focused on one or two parasite groups, while hosts are often co-infected by different taxa. The present study aims to assess the influence of environmental and host traits on the entire ectoparasite community composition of the rodent Akodon azarae. A total of 278 rodents were examined and mites (Mesostigmata), lice (Phthiraptera), ticks (Ixodida) and fleas (Siphonaptera) were determined. A multi-correspondence analysis was performed in order to analyze interactions within the ectoparasite community and the influence of environmental and host variables on this assembly. We found that environmental variables have a stronger influence on the composition of the ectoparasite community of A. azarae than the host variables analyzed. Minimum temperature was the most influential variable among the studied. In addition, we found evidence of agonistic and antagonistic interactions between ticks and mites, lice and fleas. The present study supports the hypothesis that minimum temperature plays a major role in the dynamics that shape the ectoparasite community of A. azarae, probably through both direct and indirect processes. This finding becomes particularly relevant in a climate change scenario.

Tissue-resident macrophages self-maintain locally throughout adult life with minimal contribution from circulating monocytes.

Despite accumulating evidence suggesting local self-maintenance of tissue macrophages in the steady state, the dogma remains that tissue macrophages derive from monocytes. Using parabiosis and fate-mapping approaches, we confirmed that monocytes do not show significant contribution to tissue macrophages in the steady state. Similarly, we found that after depletion of lung macrophages, the majority of repopulation occurred by stochastic cellular proliferation in situ in a macrophage colony-stimulating factor (M-Csf)- and granulocyte macrophage (GM)-CSF-dependent manner but independently of interleukin-4. We also found that after bone marrow transplantation, host macrophages retained the capacity to expand when the development of donor macrophages was compromised. Expansion of host macrophages was functional and prevented the development of alveolar proteinosis in mice transplanted with GM-Csf-receptor-deficient progenitors. Collectively, these results indicate that tissue-resident macrophages and circulating monocytes should be classified as mononuclear phagocyte lineages that are independently maintained in the steady state.

Structural organization of the bone marrow and its role in hematopoiesis.

PURPOSE OF REVIEW: The bone marrow is the main site for hematopoiesis. It contains a unique microenvironment that provides niches that support self-renewal and differentiation of hematopoietic stem cells (HSC), multipotent progenitors (MPP), and lineage committed progenitors to produce the large number of blood cells required to sustain life. The bone marrow is notoriously difficult to image; because of this the anatomy of blood cell production -- and how local signals spatially organize hematopoiesis -- are not well defined. Here we review our current understanding of the spatial organization of the mouse bone marrow with a special focus in recent advances that are transforming our understanding of this tissue. RECENT FINDINGS: Imaging studies of HSC and their interaction with candidate niches have relied on ex-vivo imaging of fixed tissue. Two recent manuscripts demonstrating live imaging of subsets of HSC in unperturbed bone marrow have revealed unexpected HSC behavior and open the door to examine HSC regulation, in situ, over time. We also discuss recent findings showing that the bone marrow contains distinct microenvironments, spatially organized, that regulate unique aspects of hematopoiesis. SUMMARY: Defining the spatial architecture of hematopoiesis in the bone marrow is indispensable to understand how this tissue ensures stepwise, balanced, differentiation to meet organism demand; for deciphering alterations to hematopoiesis during disease; and for designing organ systems for blood cell production ex vivo.

Neutrophils as regulators of the hematopoietic niche.

The niche that supports hematopoietic stem and progenitor cells (HSPCs) in the bone marrow is a highly dynamic structure. It maintains core properties of HSPCs in the steady state, and modulates their proliferation and differentiation in response to changing physiological demands or pathological insults. The dynamic and environment-sensing properties of the niche are shared by the innate immune system. Thus, it is not surprising that innate immune cells, including macrophages and neutrophils, are now recognized as important regulators of the hematopoietic niche and, ultimately, of the stem cells from which they derive. This review synthesizes emerging concepts on niche regulation by immune cells, with a particular emphasis on neutrophils. We argue that the unique developmental, circadian, and migratory properties of neutrophils underlie their critical contributions as regulators of the hematopoietic niche.

Adrenergic nerves govern circadian leukocyte recruitment to tissues.

The multistep sequence leading to leukocyte migration is thought to be locally regulated at the inflammatory site. Here, we show that broad systemic programs involving long-range signals from the sympathetic nervous system (SNS) delivered by adrenergic nerves regulate rhythmic recruitment of leukocytes in tissues. Constitutive leukocyte adhesion and migration in murine bone marrow (BM) and skeletal-muscle microvasculature fluctuated with circadian peak values at night. Migratory oscillations, altered by experimental jet lag, were implemented by perivascular SNS fibers acting on ß-adrenoreceptors expressed on nonhematopoietic cells and leading to tissue-specific, differential circadian oscillations in the expression of endothelial cell adhesion molecules and chemokines. We showed that these rhythms have physiological consequences through alteration of hematopoietic cell recruitment and overall survival in models of septic shock, sickle cell vaso-occlusion, and BM transplantation. These data provide unique insights in the leukocyte adhesion cascade and the potential for time-based therapeutics for transplantation and inflammatory diseases.

Left common iliac vein compression identified by vascular ultrasonography in asymptomatic women: does standing position influence diagnosis?

BACKGROUND: Vascular ultrasonography is the imaging exam of choice for initial screening for left common iliac vein compression, which is an asymptomatic finding that can be detected in up to 25% of some patient samples. OBJECTIVE: To determine, using vascular ultrasonography, whether findings of left common iliac vein compression in asymptomatic women are different when assessed in the prone and standing positions. METHODS: This is a cross-sectional observational study of 50 adult female volunteers with no symptoms of pelvic venous compression. The parameters assessed with vascular ultrasonography in the prone and standing positions were diameters and maximum velocities of the left common iliac vein at the point at which it crosses behind the right common iliac artery and before this point, in addition to left common iliac vein velocity indices at the crossing. RESULTS: Eight cases of significant compression of the left common iliac vein were identified when assessed in prone position (16%) and just two cases (4%) were identified in the standing position. Left common iliac vein diameters were statistically larger (p = 0.002) at the point where it crosses behind the right common iliac artery in the standing position and velocities and velocity indices were statistically higher (p < 0.001) in the prone position. No significant compression of the left common iliac vein was identified in the standing position when velocity indices were normal in the prone position. CONCLUSIONS: There was no difference in detection of significant compression of the left common iliac vein when assessed in the standing position in comparison with assessment in the prone position. However, the study showed that anatomic compression of the left common iliac vein may be reduced in the standing position.

Straightforward and Controlled Synthesis of Porphyrin-Phthalocyanine-Porphyrin Heteroleptic Triple-Decker Assemblies.

A versatile and straightforward protocol is disclosed for controlled synthesis of complex lanthanide-bridged heteroleptic porphyrin-phthalocyanine triple-decker assemblies. Two porphyrins, linked by a flexible spacer chain of intermediate length, sequentially capture lanthanide ions and a phthalocyanine to efficiently form the triple-decker complex. The bridge directs assembly, but also controls the mobility of the central macrocycle and further imparts a fully eclipsed arrangement of all three rings.