A lightweight xAI approach to cervical cancer classification.

Cervical cancer is caused in the vast majority of cases by the human papilloma virus (HPV) through sexual contact and requires a specific molecular-based analysis to be detected. As an HPV vaccine is available, the incidence of cervical cancer is up to ten times higher in areas without adequate healthcare resources. In recent years, liquid cytology has been used to overcome these shortcomings and perform mass screening. In addition, classifiers based on convolutional neural networks can be developed to help pathologists diagnose the disease. However, these systems always require the final verification of a pathologist to make a final diagnosis. For this reason, explainable AI techniques are required to highlight the most significant data to the healthcare professional, as it can be used to determine the confidence in the results and the areas of the image used for classification (allowing the professional to point out the areas he/she thinks are most important and cross-check them against those detected by the system in order to create incremental learning systems). In this work, a 4-phase optimization process is used to obtain a custom deep-learning classifier for distinguishing between 4 severity classes of cervical cancer with liquid-cytology images. The final classifier obtains an accuracy over 97% for 4 classes and 100% for 2 classes with execution times under 1 s (including the final report generation). Compared to previous works, the proposed classifier obtains better accuracy results with a lower computational cost.

Pyruvate enhances stallion sperm function in high glucose media improving overall metabolic efficiency.

If a mechanism of more efficient glycolysis depending on pyruvate is present in stallion spermatozoa, detrimental effects of higher glucose concentrations that are common in current commercial extenders could be counteracted. To test this hypothesis, spermatozoa were incubated in a 67 mM Glucose modified Tyrode's media in the presence of 1- or 10-mM pyruvate and in the Tyrode's basal media which contains 5 mM glucose. Spermatozoa incubated for 3 h at 37 °C in 67 mM Tyrode's media with 10 mM pyruvate showed increased motility in comparison with aliquots incubated in Tyrode's 5 mM glucose and Tyrode's 67 mM glucose (57.1 ± 3.5 and 58.1 ± 1.9 to 73.0 ± 1.1 %; P < 0.01). Spermatozoa incubated in Tyrode's with 67 mM glucose 10 mM pyruvate maintained the viability along the incubation (64.03 ± 15.4 vs 61.3 ± 10.2), while spermatozoa incubated in 67 mM Glucose-Tyrode's showed a decrease in viability (38.01 ± 11.2, P < 0.01). 40 mM oxamate, an inhibitor of the lactate dehydrogenase LDH, reduced sperm viability (P < 0.05, from 76 ± 5 in 67 mM Glucose/10 mM pyruvate to 68.0 ± 4.3 %, P < 0.05). Apoptotic markers increased in the presence of oxamate. (P < 0.01). UHPLC/MS/MS showed that 10 mM pyruvate increased pyruvate, lactate, ATP and NAD+ while phosphoenolpyruvate decreased. The mechanisms that explain the improvement of in presence of 10 mM pyruvate involve the conversion of lactate to pyruvate and increased NAD+ enhancing the efficiency of the glycolysis.

A Robust Ensemble of Convolutional Neural Networks for the Detection of Monkeypox Disease from Skin Images.

Monkeypox is a smallpox-like disease that was declared a global health emergency in July 2022. Because of this resemblance, it is not easy to distinguish a monkeypox rash from other similar diseases; however, due to the novelty of this disease, there are no widely used databases for this purpose with which to develop image-based classification algorithms. Therefore, three significant contributions are proposed in this work: first, the development of a publicly available dataset of monkeypox images; second, the development of a classification system based on convolutional neural networks in order to automatically distinguish monkeypox marks from those produced by other diseases; and, finally, the use of explainable AI tools for ensemble networks. For point 1, free images of monkeypox cases and other diseases have been searched in government databases and processed until we are left with only a section of the skin of the patients in each case. For point 2, various pre-trained models were used as classifiers and, in the second instance, combinations of these were used to form ensembles. And, for point 3, this is the first documented time that an explainable AI technique (like GradCAM) is applied to the results of ensemble networks. Among all the tests, the accuracy reaches 93% in the case of single pre-trained networks, and up to 98% using an ensemble of three networks (ResNet50, EfficientNetB0, and MobileNetV2). Comparing these results with previous work, a substantial improvement in classification accuracy is observed.

Non-small cell lung cancer diagnosis aid with histopathological images using Explainable Deep Learning techniques.

BACKGROUND: Lung cancer has the highest mortality rate in the world, twice as high as the second highest. On the other hand, pathologists are overworked and this is detrimental to the time spent on each patient, diagnostic turnaround time, and their success rate. OBJECTIVE: In this work, we design, implement, and evaluate a diagnostic aid system for non-small cell lung cancer detection, using Deep Learning techniques. METHODS: The classifier developed is based on Artificial Intelligence techniques, obtaining an automatic classification result between healthy, adenocarcinoma and squamous cell carcinoma, given an histopathological image from lung tissue. Moreover, a report module based on Explainable Deep Learning techniques is included and gives the pathologist information about the image's areas used to classify the sample and the confidence of belonging to each class. RESULTS: The results show a system accuracy between 97.11 and 99.69%, depending on the number of classes classified, and a value of the area under ROC curve between 99.77 and 99.94%. CONCLUSIONS: The classification results obtain a substantial improvement according to previous works. Thanks to the given report, the time spent by the pathologist and the diagnostic turnaround time can be reduced.

Seminal plasma proteins as potential biomarkers for sperm motility and velocities.

Seminal plasma proteins have important roles in sperm functionality, and different mechanisms including micro-vesicle transport of proteins are involved in the regulation of sperm biology. Due to the role of seminal plasma, we hypothesized that specific proteins present in seminal plasma may be used as discriminant variables with potential to identify stallions producing different quality ejaculates; 10 fertile stallions, with different motility and velocity values (although within normal ranges) were used in this study. Motilities and velocities were studied using computer assisted sperm analysis (CASA), while protein composition of the seminal plasma was studied using UHPLC-MS/MS. Specific proteins were more abundant in samples with poorer percentages of total motility, average path velocity and circular velocity, and were: Secreted phosphoprotein 1, Fructose-bisphosphate aldolase (p = 1,95E-09; q = 0.0005) and Malate dehydrogenase 1 (p = 1,41E-11; q = 0.002), to the contrary samples with better straight-line velocity values were enriched in Glutathione peroxidase (p=0.00013; q=0.04) and Triosephosphate isomerase (p=0.00015; q=0.04).

AnkFall-Falls, Falling Risks and Daily-Life Activities Dataset with an Ankle-Placed Accelerometer and Training Using Recurrent Neural Networks.

Falls are one of the leading causes of permanent injury and/or disability among the elderly. When these people live alone, it is convenient that a caregiver or family member visits them periodically. However, these visits do not prevent falls when the elderly person is alone. Furthermore, in exceptional circumstances, such as a pandemic, we must avoid unnecessary mobility. This is why remote monitoring systems are currently on the rise, and several commercial solutions can be found. However, current solutions use devices attached to the waist or wrist, causing discomfort in the people who wear them. The users also tend to forget to wear the devices carried in these positions. Therefore, in order to prevent these problems, the main objective of this work is designing and recollecting a new dataset about falls, falling risks and activities of daily living using an ankle-placed device obtaining a good balance between the different activity types. This dataset will be a useful tool for researchers who want to integrate the fall detector in the footwear. Thus, in this work we design the fall-detection device, study the suitable activities to be collected, collect the dataset from 21 users performing the studied activities and evaluate the quality of the collected dataset. As an additional and secondary study, we implement a simple Deep Learning classifier based on this data to prove the system's feasibility.

The SLC7A11: sperm mitochondrial function and non-canonical glutamate metabolism.

Spermatozoa are redox-regulated cells, and stallion spermatozoa, in particular, present an intense mitochondrial activity in which large amounts of reactive oxygen species (ROS) are produced. To maintain the redox potential under physiological conditions, sophisticated mechanisms ought to be present, particularly in the mitochondria. In the present study, we investigated the role of the SLC7A11 antiporter. This antiporter exchanges intracellular glutamate for extracellular cystine. In the spermatozoa, cystine is reduced to cysteine and used for GSH synthesis. The importance of the antiporter for mitochondrial functionality was studied using flow cytometry and UHPLC/MS/MS approaches. Intracellular GSH increased in the presence of cystine, but was reduced in the presence of Buthionine sulphoximine (BSO), a γ-glutamylcysteine synthetase inhibitor (P < 0.001). Inhibition of the SLC7A11 antiporter with sulfasalazine caused a dramatic drop in intracellular GSH (P < 0.001) and in the percentage of spermatozoa showing active mitochondria (P < 0.001). These findings suggest that proper functionality of this antiporter is required for the mitochondrial function of spermatozoa. We also describe that under some conditions, glutamate may be metabolized following non-conventional pathways, also contributing to sperm functionality. We provide evidences, that the stallion spermatozoa have important metabolic plasticity, and also of the relation between redox regulation and metabolic regulation. These findings may have important implications for the understanding of sperm biology and the development of new strategies for sperm conservation and treatment of male factor infertility.

Prenatal histomorphological development of the reticulum in fallow deer (Dama dama).

The histomorphological changes occurring in the Dama dama reticulum during prenatal development have been investigated. Twenty-five Dama dama embryos were used, from the first stages of prenatal life until birth. Differentiation of the reticulum was observed at 23% gestation. By 25% gestation the reticular wall comprised three layers: an internal epithelial layer, a middle layer of pluripotential blastemic tissue and an external layer or serosa. Primary reticular crests were visible at 38% gestation. Secondary reticular crests were observed at 61% gestation. Neuroendocrine cells were detected by synaptophysin (SYP) at 35% gestation, in the lamina propria-submucosa, tunica muscularis, and serosa. Epithelial Cytokeratin-18 (CK-18) cells were observed at 35% gestation extended throughout the epithelial layers. The glial cells (vimentin -VIM- and glial fibrillary acidic protein-GFAP-markers) were discerned at 25% and 43% gestation, respectively, in myenteric and submucosal plexuses, lamina propria, muscularis mucosae, tunica muscularis, and perivascular connective tissue. The neuropeptide Y (NPY) and vasoactive intestinal polypeptide (VIP) markers were immunodetected at 75% and 80 gestation, respectively, in the lamina propria-submucosa, muscularis mucosae, tunica muscularis, serosa, and myenteric plexuses. The prenatal development of the fallow deer reticular mucosa evidenced a considerable precocity similar to that previously reported in goat and red deer.

Rosiglitazone in the thawing medium improves mitochondrial function in stallion spermatozoa through regulating Akt phosphorylation and reduction of caspase 3.

BACKGROUND: The population of stallion spermatozoa that survive thawing experience compromised mitochondrial functionality and accelerated senescence, among other changes. It is known that stallion spermatozoa show very active oxidative phosphorylation that may accelerate sperm senescence through increased production of reactive oxygen species. Rosiglitazone has been proven to enhance the glycolytic capability of stallion spermatozoa maintained at ambient temperature. OBJECTIVES: Thus, we hypothesized that thawed sperm may also benefit from rosiglitazone supplementation. MATERIALS AND METHODS: Thawed sperm were washed and resuspended in Tyrodes media, and the samples were divided and supplemented with 0 or 75 µM rosiglitazone. After one and two hours of incubation, mitochondrial functionality, Akt phosphorylation and caspase 3 activity were evaluated. Additional samples were incubated in the presence of an Akt1/2 inhibitor, compound C (an AMPK inhibitor) or GW9662 (an antagonist of the PPARγ receptor). RESULTS: Rosiglitazone maintained Akt phosphorylation and reduced caspase 3 activation (p<0.01), both of which were prevented by incubation in the presence of the three inhibitors. Rosiglitazone also enhanced mitochondrial functionality (P<0.01). CONCLUSION: We provide the first evidence that the functionality of frozen stallion spermatozoa can be potentially improved after thawing through the activation of pro survival pathways, providing new clues for improving current sperm biotechnology.

The incorporation of cystine by the soluble carrier family 7 member 11 (SLC7A11) is a component of the redox regulatory mechanism in stallion spermatozoa†.

Oxidative stress is considered a major mechanism causing sperm damage during cryopreservation and storage, and underlies male factor infertility. Currently, oxidative stress is no longer believed to be caused only by the overproduction of reactive oxygen species, but rather by the deregulation of redox signaling and control mechanisms. With this concept in mind, here, we describe for the first time the presence of the soluble carrier family 7 member 11 (SLC7A11) antiporter, which exchanges extracellular cystine (Cyss) for intracellular glutamate, in stallion spermatozoa, as well as its impact on sperm function using the specific inhibitor sulfasalazine. Spermatozoa incubated with Cyss exhibited an increased intracellular GSH content compared with controls (P < 0.01): 50% in fresh extended stallion spermatozoa and 30% in frozen-thawed spermatozoa. This effect was prevented by the addition of sulfasalazine to the media. Cystine supplementation also reduced the oxidation-reduction potential of spermatozoa, with sulfasalazine only preventing this effect on fresh spermatozoa that were incubated for 3 h at 37°C, but not in frozen-thawed spermatozoa. While sulfasalazine reduced the motility of frozen-thawed spermatozoa, it increased motility in fresh samples. The present findings provide new and relevant data on the mechanism regulating the redox status of spermatozoa and suggest that a different redox regulatory mechanism exists in cryopreserved spermatozoa, thus providing new clues to improve current cryopreservation technologies and treat male factor infertility.

Comparative analysis of the merino sheep and Iberian red deer abomasum during prenatal development.

The aim of this study is to describe differences in the ontogenesis of the abomasum in sheep (domestic ruminant) and deer (wild ruminant). Histomorphometric and immunohistochemical analysis were carried out on 50 embryos and fetuses of the sheep and 50 red deer from the first prenatal stages until birth. To compare similar periods of gestation in both species, we calculate the percentages of gestation. The appearance of the abomasum was earlier in the red deer (22% gestation) than in the sheep (25% gestation). Throughout development the epithelium happened sequentially, being of the types pseudostratified to simple cylindrical. This important modification was earlier in the red deer than the sheep. At 46% gestation in red deer and 50% in sheep, gastric pits were observed on the surface of abomasal folds. Our studies suggest a close link between the initial formation of these pseudoglandular structures and the clear separation of lamina propria and submucosa separated by de muscularis mucosae. At 54% gestation in red deer and at 60% in sheep, in the bottom of these pits the first outlines of glands were distinguishable. Finally, the presence of neuroendocrine and glial cells were detected in deer at earlier stages than in sheep.

Histomorphometric study of the goat stomach during prenatal development.

The aim of this study was to perform a morphometric analysis of the different structural tissue layers of the goat stomach to study their prenatal growth from mathematical models fitted to these morphometric data. A total of 90 embryos and fetuses were used, from the early stages of prenatal life until birth. The growth rate of the gastric wall was slower than that of body length; rumen was the stomach compartment displaying slowest growth. In the three non-glandular compartments, the epithelial layer grew faster than the gastric wall itself, while the growth rate of the abomasal epithelium declined in the early stages of development. A decline in growth rate was also observed for the lamina propria and submucosa in rumen and reticulum from the early embryonic stages, whereas in omasum and abomasum these layers continued to grow as gestation progressed. The tunica muscularis displayed consistent growth in all compartments, growing faster than the gastric wall. Serosa thickness increased as gestation progressed, displaying a decline in growth-rate only in the omasum. In conclusion, the dynamics of gastric wall growth were governed by the growth rate of each of the component tissue layers.

Immunohistochemical evaluation of the goat forestomach during prenatal development.

Here we report the detection and distribution of synaptophysin (SPY), non-neuronal enolase (NNE), glial fibrillary acidic protein (GFAP), vimentin (VIM), neuropeptide Y (NPY), and vasoactive intestinal peptide (VIP) expression in the goat forestomach during prenatal development. A total of 140 embryos and fetuses were examined to evaluate protein expression from the first stage of prenatal life until birth. In all cases, SPY immunoreactivity was detected at 53 days gestation in the lamina propria-submucosa, tunica muscularis, serosa, and myenteric plexuses. Immunoreactivity to NNE was observed at 64 days gestation in the same locations as well as the epithelial layer. Glial cells were found at 64 days as indicated by signals corresponding to GFAP and VIM at 39 days. Positive staining for NPY and VIP was observed at 113, 75, and 95 days in the rumen, reticulum, and omasum, respectively, in the lamina propria-submucosa, tunica muscularis, and myenteric plexuses of each of these gastric compartments. These findings indicate possible preparation of the fetal goat forestomach for postnatal function. Compared to other ruminant species, neuroendocrine cells, glial cells and peptidergic innervations markers were detected earlier compared to sheep but at around the same stage as in deer.

Histomorphometric and immunohistochemical study of the goat reticulum during prenatal development.

This study sought to describe the morphological changes taking place in the goat reticulum during prenatal development, using histomorphometric and immunohistochemical techniques. A total of 140 goat embryos and foetuses were used, from the first stages of prenatal life until birth. Differentiation of the reticulum as a separate compartment of the primitive gastric tube was observed at 35 days of prenatal life (23% gestation). By 38 days (25% gestation) the reticular wall comprised three layers: an internal epithelial layer, a middle layer of pluripotential blastemic tissue and an external layer or serosa. Primary reticular crests were visible at 59 days (38% gestation) as evaginations of the epithelial stratum basale, marking the earliest histological differentiation of future reticular cells. Secondary reticular crests were observed at 87 days (61% gestation). Corneum papillae first became apparent on the lateral surface of primary reticular crests at 101 days (64% gestation). The muscularis mucosae was visible by 101 days (64% gestation) in primary reticular crests. Neuroendocrine cells were detected by synaptophysin at 64 days (43% gestation), while glial cell markers (glial fibrillary acidic protein and vimentin) were observed at 64 days (43% gestation) and 38 days (25% gestation), respectively. The peptidergic innervation markers such as neuropeptide Y and vasoactive intestinal polypeptide were detected at 75 days (50% gestation). In conclusion, prenatal development of the reticulum - like that of the rumen - appears to take place somewhat earlier in goats than in sheep or cattle, but at a similar rate to that reported in deer.

Histomorphometric and immunohistochemical study of the goat omasum during prenatal development.

This work studies the morphological changes taking place in the goat omasum during prenatal development, using scanning electron microscope, light microscopy and immunohistochemical analysis. A total of 140 goat embryos and fetuses were used, from the first stages of prenatal life until birth. Differentiation of the omasum as a separate compartment of the primitive gastric tube was observed at 35 days of prenatal life ([crown-rump length (CRL)] 3 cm, 23% gestation). By 38 days (CRL 4.3 cm, 25% gestation) the omasal wall comprised three layers: an internal epithelial layer, a middle layer of pluripotential blastemic tissue and an external layer or serosa. Omasal laminae appeared in the following order: primary at 38 days (CRL 4.3 cm, 25% gestation), secondary at 50 days (CRL 7.7 cm, 33% gestation), tertiary at 59 days (CRL 12 cm, 39% gestation) and quaternary at 64 days (CRL 13.5 cm, 43% gestation). Neuroendocrine cells were detected by synaptophysin (SYP) at 52 days (CRL 8 cm, 35% gestation), while glial cell markers (glial fibrillary acidic protein - GFAP, and vimentin-VIM) were observed at 64 days (CRL 13.5 cm, 43% gestation) and 38 days (CRL 4.3 cm, 25% gestation), respectively. Sympathetic and parasympathetic nerve fibers and nerve bodies were detected via neuropeptide Y (NPY) and vasoactive intestinal polypeptide (VIP) at 95 days (CRL 20 cm, 63% gestation). In conclusion, prenatal development of the omasum - like that of the rumen - appears to take place somewhat earlier in goats than in sheep or cattle, but at a similar stage to that reported in deer.

Histomorphometric and immunohistochemical study of the goat rumen during prenatal development.

This study sought to chart the ontogenesis of the goat rumen by histomorphometric examination, scanning electron microscopy and immunohistochemical analysis. A total of 140 goat embryos and fetuses were used, from the first stage of prenatal life until birth. The appearance of the rumen from the primitive gastric tube was observed at 35 days of prenatal life (CRL 3 cm, 23% gestation). By 38 days (CRL 4.3 cm CRL, 25% gestation) the ruminal wall comprised three layers: an internal epithelial layer, a middle layer of pluripotential blastemic tissue and an external layer or serosa. Ruminal pillars were visible at 46 days (CRL 6 cm, 30% gestation), and by 76 days (CRL 18 cm, 50% gestation) ruminal papillae were starting to appear. Under scanning electron microscopy, by 50 days (CRL 7.7 cm, 33% gestation) small ruminal papillae were observed protruding from the surface. Finally, neuroendocrine cells (synaptophysin, SYP) were detected at 53 days (CRL 9 cm CRL, 35% gestation), while glial cell markers (glial fibrillary acidic protein--GFAP, and vimentin-VIM) were found at 108 days (CRL 31 cm, 72% gestation) and 39 days (CRL 4.4 cm, 26% gestation), respectively. Neuropeptide Y (NPY) and vasoactive intestinal polypeptide (VIP) were detected immunohistochemically at 113 days (CRL 33 cm, 75% gestation) and 120 days (CRL 35 cm, 80% gestation), respectively. In conclusion, histomorphogenesis of the rumen in goats was similar to that reported in deer, but rather slower than observed for sheep or cattle.

Ontogenesis of the rumen: a comparative analysis of the Merino sheep and Iberian red deer.

The aim of this study is to describe differences in the ontogenesis of the rumen in the sheep (domestic ruminant) and deer (wild ruminant). A total of 50 embryos and fetuses of Merino sheep and 50 of Iberian deer were used, from the first stages of prenatal life until birth. For the study, the animals were divided into five experimental groups according to the most relevant histological characteristics. The appearance of the rumen from the primitive gastric tube was earlier in the sheep (22% gestation, 33 days) than in the deer (25% gestation, 66 days). In both cases it displayed a primitive epithelium of a stratified, cylindrical, non-ciliary type. At around 28% gestation in the sheep (42 days) and 26% (67 days) in the deer, the rumen was configured of three clearly-differentiated layers: internal or mucosal, middle or muscular and external or serosal. In both species the stratification of the epithelial layer was accompanied by modifications in its structure with the appearance of the ruminal pillars and papillae. The pillars appeared before the papillae and the appearance of both structures was always earlier in the deer (pillars: 70 days, 27% gestation; papillae: 97 days, 36% gestation) than in the sheep (pillars: 42 days, 28% gestation; papillae: 57 days, 38% gestation). The outlines of the ruminal papillae appeared as evaginations of the basal zone toward the ruminal lumen, dragging in their formation the basal membrane, the lamina propria and the submucosa. The tegumentary mucosa of the rumen was without secretion capability in the first embryonic phases. From 67 days (26% gestation) the neutral mucopolysaccharides appeared in the deer and at 46 days (30% gestation) in the sheep. In both cases they continued to decrease until birth, this diminution being more pronounced in the deer. Finally, the presence of neuroendocrine and glial cells was detected in the deer at earlier stages than in the sheep.