Entire Organism Research Articles

Identification of regional variation in gene expression and inflammatory proteins in donor lung tissue and ex vivo lung perfusate

ObjectiveDiagnosing lung injury is a challenge in lung transplantation. It has been unclear if a single biopsy specimen is truly representative of the entire organ. Our objective was to investigate lung inflammatory biomarkers using human lung tissue biopsies and ex vivo lung perfusion perfusate. MethodsEight human donor lungs declined for transplantation were air inflated, flash frozen, and partitioned from apex to base. Biopsies were then sampled throughout the lung. Perfusate was sampled from 4 lung lobes in 8 additional donor lungs subjected to ex vivo lung perfusion. The levels of interleukin-6, interleukin-8, interleukin-10, and interleukin-1β were measured using quantitative reverse transcription polymerase chain reaction from lung biopsies and enzyme-linked immunosorbent assay from ex vivo lung perfusion perfusate. ResultsThe median intra-biopsy equal-variance P value was .50 for messenger RNA biomarkers in tissue biopsies. The median intra-biopsy coefficient of variance was 18%. In donors with no apparent focal injuries, the biopsies in each donor showed no difference in various lung slices, with a coefficient of variance of 20%. The exception was biopsies from the lingula and injured focal areas that demonstrated larger differences. Cytokines in ex vivo lung perfusion perfusate showed minimal variation among different lobes (coefficient of variance = 4.9%). ConclusionsCytokine gene expression in lung biopsies was consistent, and the biopsy analysis reflects the whole lung, except when specimens were collected from the lingula or an area of focal injury. Ex vivo lung perfusion perfusate also provides a representative measurement of lung inflammation from the draining lobe. These results will reassure clinicians that a lung biopsy or an ex vivo lung perfusion perfusate sample can be used to inform donor lung selection.

Highly specific and non-invasive imaging of Piezo1-dependent activity across scales using GenEPi

Mechanosensing is a ubiquitous process to translate external mechanical stimuli into biological responses. Piezo1 ion channels are directly gated by mechanical forces and play an essential role in cellular mechanotransduction. However, readouts of Piezo1 activity are mainly examined by invasive or indirect techniques, such as electrophysiological analyses and cytosolic calcium imaging. Here, we introduce GenEPi, a genetically-encoded fluorescent reporter for non-invasive optical monitoring of Piezo1-dependent activity. We demonstrate that GenEPi has high spatiotemporal resolution for Piezo1-dependent stimuli from the single-cell level to that of the entire organism. GenEPi reveals transient, local mechanical stimuli in the plasma membrane of single cells, resolves repetitive contraction-triggered stimulation of beating cardiomyocytes within microtissues, and allows for robust and reliable monitoring of Piezo1-dependent activity in vivo. GenEPi will enable non-invasive optical monitoring of Piezo1 activity in mechanochemical feedback loops during development, homeostatic regulation, and disease.

Eroticism as a hormetic stimulus in health and ageing

Eroticism in later life is, on the whole, a taboo subject, and the stigma attached to expressions of sexual intent by older people is widespread in most cultures. However, sexuality and eroticism have an important role to play in maintaining healthy ageing. Sexuality is an essential aspect of our biology and its effects have repercussions in systems and organs other than the sexual. In this paper I review the importance of developing a sexual-erotic element in later life, an element that is intimately coupled with the phenomenon of hormesis. In hormesis, there is biological benefit following exposure to a mild stimulus or challenge, whereas exposure to a higher dose of the same stimulus becomes detrimental. Innovative sexual stimulation can be seen as a hormetic opportunity to initiate beneficial effects on the entire ageing human organism. The intention is to show that, health benefits may be obtained not only through physical (exercise), nutritional, mental or pharmacological challenges, but also through sexual stimulation of the appropriate magnitude and quality. By enhancing erotic stimulation it may be possible to experience many other benefits spanning several domains such as the endocrine, immune, circulatory and neurological. In addition, within an increasingly information-rich world, sexuality may play a part in improving adaptation to various cognitive external stimuli originating from our technological ecosystem.

Fecal microbiota transplantation: A new strategy to delay aging

The gut microbiota, the community of gut microorganisms that inhabit the gastrointestinal tract, plays a crucial role in modulating host immunity, metabolism, and neurological health, thereby influencing the entire organism. Aging is associated with changes in gut microbiota composition and functionality, often resulting in dysbiosis, an imbalance of gut microbiota and an aging hallmark. Recent studies have suggested that fecal microbiota transplantation (FMT), the transfer of fecal material containing beneficial microbes from a healthy donor to a recipient with a disturbed microbiota, may restore the balance of host gut microbiota and ameliorate some aging-associated impairments in diverse organs. In particular, FMT from young donors has shown more beneficial effects than FMT from aged donors. In this paper, we review recent advances in FMT for its effects on aging and discuss the potential mechanisms and challenges of this novel intervention, highlighting its potential to improve health outcomes in the aging population.

An Ayurvedic & Contemporary Perspective on a Comprehensive Appraisal of Ageing

As well as being a Swabhavaja Vyadhi (natural sickness), Avurveda has seen Jara or Vardhakya as a natural and unavoidable process. When an organ of a different system begins to degenerated and lose its anatomical and physiological capabilities over time, it is said to be ageing. This process can occur in a cell, an organ, or the entire organism.
 Shareera (physical), Indriya (emotional), Satwa (psychological level), Agni (metabolism), Bala and Ojas etc. are influencing factors for Jara (ageing). The final stage of life is called Vriddhavastha (Jara). During this phase, every aspect of the body begins to degenerate, including the Dhatu (different anatomical tissues), the Indriya (sensory and motor organ), the ability to perceive, the power of speech, the power of the body, and numerous mental and cognitive functions (such as memory, intellect, reception, retention, analytic ability, etc).
 The Vayu dosha predominates throughout this phase (one of the three physiological body factors). The most noticeable physical changes at this time include ageing skin, greying hair, baldness, and a decline in physical stamina. Loss of muscle compactness, joint looseness, vitiation of Rakta (blood), excessive production of Meda (fatty tissue), failure of Majja (marrow) accumulation in bone, failure of production of Shukra (semen), and loss of Ojas component are examples of these pathological alterations. This article is made to explore the physiological changes during Aging.

Stigmaria: on the substrate before in the substrate

Stigmaria is the rooting organ of late Paleozoic arborescent lycopsids. Previous scientific considerations of the stigmarian habit have placed the entire rooting organ shallow beneath the substrate surface, despite the proximal main stigmarian axes radiating outwards orthogonally from the trunk in all known stump casts with attached Stigmaria. Here, we show definitively that stigmarian axes were subaerial proximally given both their preservation within and direct interaction with two independent volcanic ash layers. However, it remains uncertain whether these axes dove under the substrate surface distally and, if so, where along the axes this occurred. We test the aboveground or belowground nature of distal stigmarian axes with comparisons of sampling frequencies of vasculature from different regions of the arborescent lycopsid body plan as preserved in ‘coal ball’ permineralization fossils from the Pennsylvanian of Euramerica. The overabundance of root versus shoot material has previously been recognized as evidence of the enhanced preservation potential of organs growing within, rather than above, the substrate. We find that specifically the terminal stigmarian axis vasculature is overrepresented in the fossil record compared to trunk and proximal stigmarian axis vasculature, suggesting that the primary stigmarian axes and their secondary branches grew on the substrate before eventually going underground distally. Implications for arborescent lycopsid establishment are considered.

Four Components of the Mobilization of the Russian Sciences

The transition of Russian science to the mobilization scenario of development is considered. The survey data are presented, demonstrating that this transition causes a positive attitude among the majority of the Russian scientific community. The author explains this rather paradoxical fact with two reasons. Firstly, by the fact that scientists are tired of the Russian science being in a state of Russian “nirvana”, when work in the “not shaky, not shaky” mode, at a sufficiently low level of its capabilities, has been raised to normal. Secondly, they understand that in the current conditions, Russia is mainly rescued by the presence of nuclear weapons created during the mobilization of domestic science. The mobilization scenario of the development of Russian science is considered as including four main components: 1) stricter control (by the state) of the issues being developed, 2) optimization of the staff of science, 3) change in the (meritocratic) form of its organization, 4) intensification of scientific work. The question is raised about which part of the national science it is advisable to mobilize. The author believes that the mobilization of our entire scientific organism would be an excess, but some of its elements – the liberation from “dead souls”, the intensification of scientific work, etc., should be carried out in relation to our entire science. The risks facing Russian science in the conditions of its mobilization are also considered: de-internationalization, reduction of international investments, etc.

Phytohormonal balance in leaves of the soft wheat lines isogenic for VRN genes

The paper presents results of the study of the phytohormonal balance in mature, formed leaves of the lines of common wheat (Triticum aestivum L.) of the winter variety Olvia during the transition from vegetative to generative development. Near isogenic for the VRN genes lines (NILs) created in the gene pool of the Olvia variety and plants of the vernalized and non-vernalized winter variety were used. The experiments were carried out under the conditions of a vegetation experiment in the factorial chamber of the Department of Physiology and Biochemistry of Plants and Microorganisms of the V.N. Karazin KhNU. During the experiment, we conducted physiological observations and analyzed the development rate of experimental plants. Phytohormone analysis was carried out in fixed plant material by a chromatographic distribution of phytohormone mixture with thin-layer chromatography. The phytohormones were identified by the reference standards irradiating the chromatograms with ultraviolet UV (254 nm), and the content was determined by biotesting methods. The level of main classes of classical growth-stimulating phytohormones (auxins (IAA), cytokinins (CK), and gibberellins (GA), and growth-inhibiting hormones (abscisins (ABA)) was analyzed. The indicators of phytohormonal balance were calculated as the ratio of growth-stimulating and growth-inhibiting hormones. The results of the experiments showed that phytohormones in mature, formed leaves of the experimental plants are represented by auxins – 64.9-70.7 μg/g, cytokinins – 26.6-30.5 μg/g, gibberellins – 179.47-228.68 μg/g, and abscisins – 54.06-89.76 μg/g of dry weight. Among the phytohormone classes studied, the minimal was the cytokinins’ content, while the phytohormones of terpenoid nature (gibberellins and abscisins) were represented best. It has been established that the phytohormonal balance viz. the ratio of growth-stimulating and growth-inhibiting phytohormones reflects the development rate of experimental plants. Rapidly developing plants of isolines VRN 1 and VRN 3, and the plants of vernalized variety were characterized by the maximum phytohormone balance (especially GA/ABA), while the slowly developing plants of the isoline VRN 2 and the non-vernalized variety Olvia had the minimum balance. Since the plant organism is an integrated system of organs and functions, we assume that this indicator – phytohormonal balance in plant leaves, can be used as a marker of the ontogenetic state of the entire plant organism. The identified changes in the phytohormonal status of mature, formed leaves and the development rates of experimental plants have the same regularities in all the models used in our research: the model of isogenic lines and the model of vernalized and non-vernalized plants of the winter variety. This fact makes it possible to assume that changes in the phytohormone balance of mature leaves, which reflect the ontogenetic state of the entire plant organism, are determined by the genotypic and phenotypic (epigenetic) influence.

Nanotechnology-Based Vaccines

Several new infectious diseases have developed in recent years, and a few old ones that were formerly thought to pose no threat to humans have made a comeback. Millions of fatalities are attributed to these illnesses together, having a significant negative influence on the worldwide socioeconomic and healthcare sectors. The lack of appropriate medications for many of these disorders is one of the biggest obstacles to treating them. Yet, several of the most common diseases currently have no vaccinations that are reliable. The ideal vaccine should have several key characteristics, including safety, stability, and the capacity to induce a sufficient and long-lasting immune response with a minimal number of doses. To induce protective immunity against illnesses, different generation vaccines are employed, including attenuated or dead entire organisms (first generation), subunits (second generation), and RNA or DNA vaccines (third generation). To get beyond these obstacles, a reliable vaccination delivery mechanism is needed, one that not only gets the vaccine molecules to the target region where they can trigger long-lasting immune responses but also has few side effects and uses fewer doses. Only a few hundred atoms make up the majority of nanoparticles. Nanoparticles have a relatively significant surface area-to-volume ratio because of their extremely small size. Nanoparticles can have surprising optical, physical, and chemical properties due to this property. Nanotechnology has many benefits for the creation of vaccines for the next generation. A delivery strategy based on nanocarriers can shield vaccines from early deterioration, increase stability, have high adjuvant qualities, and can help with the targeted distribution of an immunogen. The researcher conducts an examination of articles that are in accordance with the issue to be studied. Articles used in the literature review are obtained through the database of international journal providers through PubMed, we investigated clinical studies and discussed what happened in these clinical studies and the extent of the effectiveness of Nanoparticle-Based Vaccines. In order to achieve effective vaccine distribution and generate the required host immunity against infectious diseases, this review article focuses on the applications of nanocarrier-based vaccine formulations and the methodologies utilized for functionalizing nanoparticles.

NISNet3D: three-dimensional nuclear synthesis and instance segmentation for fluorescence microscopy images

The primary step in tissue cytometry is the automated distinction of individual cells (segmentation). Since cell borders are seldom labeled, cells are generally segmented by their nuclei. While tools have been developed for segmenting nuclei in two dimensions, segmentation of nuclei in three-dimensional volumes remains a challenging task. The lack of effective methods for three-dimensional segmentation represents a bottleneck in the realization of the potential of tissue cytometry, particularly as methods of tissue clearing present the opportunity to characterize entire organs. Methods based on deep learning have shown enormous promise, but their implementation is hampered by the need for large amounts of manually annotated training data. In this paper, we describe 3D Nuclei Instance Segmentation Network (NISNet3D) that directly segments 3D volumes through the use of a modified 3D U-Net, 3D marker-controlled watershed transform, and a nuclei instance segmentation system for separating touching nuclei. NISNet3D is unique in that it provides accurate segmentation of even challenging image volumes using a network trained on large amounts of synthetic nuclei derived from relatively few annotated volumes, or on synthetic data obtained without annotated volumes. We present a quantitative comparison of results obtained from NISNet3D with results obtained from a variety of existing nuclei segmentation techniques. We also examine the performance of the methods when no ground truth is available and only synthetic volumes were used for training.

Transcriptome analysis reveals an Atoh1b-dependent gene set downstream of Dlx3b/4b during early inner ear development in zebrafish.

The vertebrate inner ear is the sensory organ mediating hearing and balance. The entire organ develops from the otic placode, which itself originates from the otic-epibranchial progenitor domain (OEPD). Multiple studies in various species have shown the importance of the forkhead-box and distal-less homeodomain transcription factor families for OEPD and subsequent otic placode formation. However, the transcriptional networks downstream of these factors are only beginning to be understood. Using transcriptome analysis, we here reveal numerous genes regulated by the distal-less homeodomain transcription factors Dlx3b and Dlx4b (Dlx3b/4b). We identify known and novel transcripts displaying widespread OEPD expression in a Dlx3b/4b-dependent manner. Some genes, with a known OEPD expression in other vertebrate species, might be members of a presumptive vertebrate core module required for proper otic development. Moreover, we identify genes controlling early-born sensory hair cell formation as well as regulating biomineral tissue development, both consistent with defective sensory hair cell and otolith formation observed in dlx3b/4b mutants. Finally, we show that ectopic Atoh1b expression can rescue early sensorigenesis even in the absence of Dlx3b/4b. Taken together, our data will help to unravel the gene regulatory network underlying early inner ear development and provide insights into the molecular control of vertebrate inner ear formation to restore hearing loss in humans ultimately.

In situ hybridization analysis of olfactory receptor expression in the sea turtle olfactory organ.

The olfactory organ of turtles consists of an upper chamber epithelium (UCE) with associated glands, and a lower chamber epithelium (LCE) devoid of glands. The UCE and LCE are referred to as the air-nose and the water-nose, respectively, because the UCE is thought to detect airborne odorants, while the LCE detects waterborne odorants. However, it is not clear how the two are used in the olfactory organ. Odorant receptors (ORs) are the major olfactory receptors in turtles; they are classified as class I and II ORs, distinguished by their primary structure. Class I ORs are suggested to be receptive to water-soluble ligands and class II ORs to volatile ligands. This study analyzed the expression of class I and II ORs in hatchlings of the green sea turtle, Chelonia mydas, through in situ hybridization, to determine the localization of OR-expressing cells in the olfactory organ. Class I OR-expressing cells were distributed mainly in the LCE, implying that the LCE is receptive to waterborne odorants. Class II OR-expressing cells were distributed in both the UCE and LCE, implying that the entire olfactory organ is receptive to airborne odorants. The widespread expression of class II ORs may increase opportunities for sea turtles to sense airborne odorants.

Acute radiation syndrome

Over the last decades, along with progress and development of using ionizing radiation, including the fields of power engineering or nuclear weapons, it was possible to observe and describe negative effects of the radiation on human body. The phenomenon referred to as acute radiation syndrome describes the effects of radiation on living cells, depending on exposure time and absorbed dose, which affects the functioning of entire organism after the exposure. The following work describes how ionizing radiation causes changes at the cellular and tissue level, both directly and indirectly, taking into account the variables affecting final effects of exposure. Consecutive phases of acute radiation syndrome were distinguished, and the focus was also on individual groups of disorders resulting from exposure, which may overlap and coexist, i.e. hematopoietic syndrome, gastrointestinal syndrome and neurovascular syndrome, as well as developing multi-organ failure in some of the cases, leading to the patient's death. Exposure to ionizing radiation can result in diversified, variably intensified and spread over time symptoms from many organ systems. Depending on the characteristics of the exposure, its effects will differ, and thus the prognosis for the patient.

“Autophagic landscapes: on the paradox of survival through self-degradation” – a science-inspired exhibition

ABSTRACT The Complexity Science Hub Vienna is hosting an autophagy-based art exhibition that shows the artwork by Ayelen Valko and Dorotea Fracchiolla, two artists who are also scientists engaged in autophagy research. This exhibition, called “Autophagic landscapes: on the paradox of survival through self-degradation”—which will be open to the general public from January to May 2023—proposes a visual journey from entire organisms toward the interior of a single cell. The core ideas represented in the exhibited artworks are the molecular mechanisms and vesicular dynamics of autophagy-two phenomena that have been feeding the imagination of the two artists, inspiring the creation of art that depicts intriguing subcellular landscapes. Although the microscale bears very valuable aesthetic features, it is not a common subject in art. Correcting this is the main aim of this exhibition and of the two artists.

Antimicrobial and Antioxidant Evaluation of Various Parts of Gossypium Herbacium (Linn) Plant

The methanol extracts of the various plant parts were screened for anti-oxidant activity by thin layer chromatography using 2,2-diphenyl-1-dipicrylhydrazyl (DPPH) while in vitro antimicrobial evaluation was determined by agar diffusion method and bioautographic technique using Escherichia coli, Bacillus subtilis, Staphylococcus aureus and Pseudomonas aeruginosa as test organisms; aqueous solvent served as control and ciprofloxacin served as standard drug. The result showed that methanol extracts of the stem, leaf, root and seed of the plant were found to be stronger against the entire test organisms and the stem and leaf of the plant were also found to possess strong antioxidant activity. The antimicrobial property shown by the leaf is an evidence of the ethno medicinal uses of the plant. G. herbaceum leaf, seed, epicarp, stem bark, root bark, may provide novel plant-derived therapeutic agents, effective in treating infectious diseases arising from multiple drug-resistant bacteria and a target in the management of oxidative stress that could easily lead to aging.

The Influence of Gut Microbiota on Oxidative Stress and the Immune System.

The human gastrointestinal tract is home to a complex microbial community that plays an important role in the general well-being of the entire organism. The gut microbiota generates a variety of metabolites and thereby regulates many biological processes, such as the regulation of the immune system. In the gut, bacteria are in direct contact with the host. The major challenge here is to prevent unwanted inflammatory reactions on one hand and on the other hand to ensure that the immune system can be activated when pathogens invade. Here the REDOX equilibrium is of utmost importance. This REDOX equilibrium is controlled by the microbiota either directly or indirectly via bacterial-derived metabolites. A balanced microbiome sorts for a stable REDOX balance, whereas dysbiosis destabilizes this equilibrium. An imbalanced REDOX status directly affects the immune system by disrupting intracellular signaling and promoting inflammatory responses. Here we (i) focus on the most common reactive oxygen species (ROS) and (ii) define the transition from a balanced REDOX state to oxidative stress. Further, we (iii) describe the role of ROS in regulating the immune system and inflammatory responses. Thereafter, we (iv) examine the influence of microbiota on REDOX homeostasis and how shifts in pro- and anti-oxidative cellular conditions can suppress or promote immune responses or inflammation.

Low Exercise Tolerance as a Marker of Erectile Dysfunction and Depression among Post-Myocardial Infarction Men.

Evidence has grown recently on the correlation between lifestyle and physical activity, and their impact on the functioning of the entire organism. In addition, a decrease in physical efficiency may be an indicator of the early diagnosis of systemic diseases. The aim of this study was to determine whether there is a relationship between exercise tolerance and possible erectile dysfunction or mental disorders. A cross-sectional study was conducted among 254 men in 4 cardiac rehabilitation centers in Poland using the standardized International Index of Erectile Function 5 (IIEF-5) and Beck Depression Inventory (BDI) questionnaires. Erectile dysfunction was directly proportional to the metabolic equivalent of the task (MET) variable. An increase in exercise tolerance by 1 point was associated with an increase in the IIEF-5 score by 1.62 points, indicating a reduction in the severity of erectile dysfunction. The 1-point increase in MET was associated with a 1.8-point decrease in BDI, indicating that an increase in exercise tolerance is associated with a decrease in the severity of depressive disorders. Increasing the tolerance of physical effort has a beneficial effect not only on the general well-being, but also on the sexual and mental health of men. An increase in exercise tolerance is associated with a reduction in the incidence of erectile dysfunction. On the other hand, in patients with depressive disorders, the improvement of exercise tolerance reduces the intensity of depression symptoms. Exercise tolerance can be an early and simple marker at the onset of erectile dysfunction or mood disorders.

Cellular and molecular effects of non-ionizing electromagnetic fields.

The way that living cells respond to non-ionizing electromagnetic fields (EMF), including static/extremely-low frequency and radiofrequency electromagnetic fields, fits the pattern of 'cellular stress response'- a mechanism manifest at the cellular level intended to preserve the entire organism. It is a set pattern of cellular and molecular responses to environmental stressors, such as heat, ionizing radiation, oxidation, etc. It is triggered by cellular macromolecular damage (in proteins, lipids, and DNA) with the goal of repairing and returning cell functions to homeostasis. The pattern is independent of the type of stressor encountered. It involves cell cycle arrest, induction of specific molecular mechanisms for repair, damage removal, cell proliferation, and cell death if damage is too great. This response could be triggered by EMF-induced alternation in oxidative processes in cells. The concept that biological response to EMF is a 'cellular stress response' explains many observed effects of EMF, such as nonlinear dose- and time-dependency, increased and decreased risks of cancer and neurodegenerative diseases, enhanced nerve regeneration, and bone healing. These responses could be either detrimental or beneficial to health, depending on the duration and intensity of the exposure, as well as specific aspects ofthe living organism being exposed. A corollary to electromagnetic hypersensitivity syndrome (EHS) could be an inappropriate response of the hippocampus/limbic system to EMF, involving glucocorticoids on the hypothalamic-pituitary-adrenal axis.

Pragmatic applications of DNA barcoding markers in identification of fish species – A review

Abstract DNA barcoding and mini-barcoding involve Cytochrome Oxidase Subunit I (COI) gene in mitochondrial genome and are used for accurate identification of species and biodiversity. The basic goal of the current study is to develop a complete reference database of fishes. It also evaluates the applicability of COI gene to identify fish at the species level with other aspects i.e., as Kimura 2 parameter (K2P) distance. The mean observed length of the sequence was ranging from 500 to 700 base pairs for fish species in DNA barcoding and from 80 to 650 base pairs for DNA mini-barcoding. This method describes the status of known to unknown samples but it also facilitates the detection of previously un-sampled species at distinct level. So, mini-barcoding is a method which focuses on the analysis of short-length DNA markers and has been demonstrated to be effective for species identification of processed food containing degraded DNA. While DNA metabarcoding refers to the automated identification of multiple species from a single bulk sample. They may contain entire organisms or a single environmental sample containing degraded DNA. Despite DNA barcoding, mini-barcoding and metabarcoding are efficient methods for species identification which are helpful in conservation and proper management of biodiversity. It aids researchers to take an account of genetic as well as evolutionary relationships by collecting their morphological, distributional and molecular data. Overall, this paper discusses DNA barcoding technology and how it has been used to various fish species, as well as its universality, adaptability, and novel approach to DNA-based species identification.

COMBINe enables automated detection and classification of neurons and astrocytes in tissue-cleared mouse brains

Tissue clearing renders entire organs transparent to accelerate whole-tissue imaging; for example, with light-sheet fluorescence microscopy. Yet, challenges remain in analyzing the large resulting 3D datasets that consist of terabytes of images and information on millions of labeled cells. Previous work has established pipelines for automated analysis of tissue-cleared mouse brains, but the focus there was on single-color channels and/or detection of nuclear localized signals in relatively low-resolution images. Here, we present an automated workflow (COMBINe, Cell detectiOn in Mouse BraIN) to map sparsely labeled neurons and astrocytes in genetically distinct mouse forebrains using mosaic analysis with double markers (MADM). COMBINe blends modules from multiple pipelines with RetinaNet at its core. We quantitatively analyzed the regional and subregional effects of MADM-based deletion of the epidermal growth factor receptor (EGFR) on neuronal and astrocyte populations in the mouse forebrain.