Evaluation of the morphology and development of preantral ovarian follicles in mice submitted to a chronic diet of dietary supplementation with Pereskia aculeata Miller leaves.

This study aimed to investigate the effect of including mouse feed with different concentrations (5, 10, or 20%) of Pereskia aculeata Miller (PAM) leaves on the morphology and development of preantral ovarian follicles and ovarian stromal cell density. The oral toxicity was performed using repeated dose toxicity assays subdivided into experiments of 30 days and 90 days of treatment. After the experiments, the ovaries of each animal were collected and submitted to classical histology. At 30 and 90 days, there was an equivalent percentage of normal, primordial, and developing follicles (P > 0.05) between PAM treatments compared to the control. Regarding the different stages of follicular development, after 90 days, there was a higher percentage (P < 0.05) of developing follicles only in the control group compared to day 30. The PAM 5% treatment was the only one that affected the cell density in the stroma after 90 days of treatment. Thus, we observed that supplementing the diet with P. aculeata did not pose any risk concerning animal consumption; specifically, there were no toxic reproductive effects observed from adding Pereskia aculeata Miller to the mouse diet.

Dataset of the effect of the number of injector holes on the heat transfer coefficient and the pressure in the combustion chamber of a hydrogen-diesel engine.

There are several methods for simulating internal combustion engines. The computational fluid dynamics method is the best way to simulate these engines because it can simulate the combustion process, which is a microscopic process. In this study, the simulation of the combustion process in a closed cycle in a diesel engine with a mixture of diesel and hydrogen is done by AVL Fire software. In order to simulate the combustion in the Species and chemical transmission section, a chemkin mechanism is coupled with AVL Fire software. In this study, the effect of 10 % hydrogen fuel and 90 % diesel fuel as well as the effect of nozzle holes (1, 3 and 6 holes) on the engine performance were directly investigated. In order to validate the results of the pressure simulation and the temperature inside the cylinder in the diesel fuel combustion mode, at 2800 rpm and 100 % load, the data were compared with the experimental data. The research also included verification of the heat transfer coefficient (HTC) results with theoretical data obtained by Woschni and Hohenberg. To accurately simulate the combustion process, the simulation data was validated by comparing the pressure and temperature inside the cylinder at a specific operating condition with experimental data. The results indicate that the maximum heat transfer coefficient is achieved at the angle of maximum pressure, with the exhaust valve having the highest coefficient. The addition of hydrogen to diesel fuel results in a 1.72 % increase in the heat transfer coefficient due to increased collisions. In addition, the introduction of hydrogen fuel increases cylinder pressure and engine power, while increasing the number of fuel nozzle holes decreases the coefficient and pressure, which affects fuel penetration and evaporation rate.

Correction: Bufalo et al. Human Sensory Neuron-like Cells and Glycated Collagen Matrix as a Model for the Screening of Analgesic Compounds. Cells 2022, 11, 247.

In the original publication [...].

Effects of high-intensity interval training on subjective sleep quality and daytime sleepiness in patients with obstructive sleep apnea: A secondary analysis from a randomized controlled trial.

This study aimed to verify the effect of 12 weeks of HIIT on the perceived sleep quality and excessive daytime sleepiness in patients with obstructive sleep apnea (OSA). For this, a secondary analysis of a randomized controlled trial, including 36 adults with moderate-severe OSA (19 males; 52.2 ± 9.8 years; body mass index = 34.2 ± 5.8; AHI = 42.0 ± 22.9 e/h) was performed. Participants were randomly assigned to HIIT [5 periods of 4 min of walking or running on a treadmill at 90-95 % of maximum heart rate (HRmax) interspersed with 3 min of walking at 50-55 % of HRmax performed three times per week for 12 weeks] or a control group (CG; stretching exercises performed two times per week for 12 weeks). Specific domains of subjective sleep quality and EDS were assessed at baseline and post 12 weeks. Generalized estimated equation were used to verify between groups and times differences. There were no group × time interactions for the domains sleep duration (0.416), sleep efficiency (0.198), sleep disturbance (0.523), and sleep medications (0.915). However, significant group × time interactions were observed for global sleep score (0.022), and for the domains sleep quality (0.001), sleep latency (0.029), and daytime dysfunction (0.012). In addition, there was a significant group × time interaction for EDS (HIIT = -3.4 ± 0.9; CG change = -1.0 ± 1.0; p = 0.023). Thus, in patients with OSA, 12 weeks of HIIT improves perceived sleep quality and daytime sleepiness.

White wine vinification and an expanded insight into pellicular macerations: bibliometric and bibliographic review.

BACKGROUND: Pellicular macerations in the vinification of white wines involve the contact of grape skins and seeds with the must before, during or after alcoholic fermentation. Pre-fermentative pellicular maceration aims to enrich the must with volatile compounds and aroma precursors. Fermentative maceration occurs during alcoholic fermentation, whereas post-fermentative maceration is carried out after this process, associated with orange, amber or skin-contact wines, which have experienced a growing global demand in recent years. In this context, this research aimed to conduct a bibliometric review on pellicular macerations in white wines using two search strategies on the specific platform for the period from 2010 to 2023. Additionally, we sought to identify research trends in this segment of the wine industry through a comprehensive literature review of the retrieved documents. RESULTS: The results emphasized more studies on pre-fermentative pellicular maceration than on long-duration macerations during and after alcoholic fermentation. Alternative maceration techniques, such as grape freezing, were also observed as study subjects, including their effects on final wines. The research identified a wide variety of grapes explored in studies related to pellicular macerations of the Vitis vinifera L. species, with approximately 50 distinct nomenclatures identified. Regarding pre-fermentative macerations, the contact time varied from 2 to 60 h, with the temperature range 1-20 °C. CONCLUSION: The specific search for extended skin contact white wines revealed a limited number of available documents, indicating that studies related to this product style are promising and necessary, given the growing commercial relevance of this product profile. © 2024 Society of Chemical Industry.

ACTH-producing adrenocortical carcinoma: an exceedingly rare diagnosis.

BACKGROUND: Adrenocortical carcinoma is a very rare endocrinopathy that has a poor prognosis and is frequently associated with ACTH-independent Cushing's syndrome. Despite having an adrenocortical carcinoma, our patient surprisingly had an ACTH-dependent Cushing's syndrome. CASE REPORT: A 26-year-old female presented with Cushing's syndrome and an abdominal mass. Imaging studies revealed an adrenal mass consistent with a high-grade malignancy. Laboratory workup showed hypercortisolism, hyperandrogenism, and hypokalemia with normal levels of metanephrines. Unexpectedly, her ACTH levels were remarkably elevated. The pathological analysis of a tumor sample was conclusive for adrenocortical carcinoma with immunopositivity for ACTH. CONCLUSIONS: Our patient suffered from an adrenocortical carcinoma that was ectopically producing ACTH. This case emphasizes that physicians should have a broad-minded approach when evaluating cases of rare endocrine malignancies.

Time Scale Calculus: a new approach to multi-dose pharmacokinetic modeling.

In this paper, we use Time Scale Calculus (TSC) to formulate and solve pharmacokinetic models exploring multiple dose dynamics. TSC is a mathematical framework that allows the modeling of dynamical systems comprising continuous and discrete processes. This characteristic makes TSC particularly suited for multi-dose pharmacokinetic problems, which inherently feature a blend of continuous processes (such as absorption, metabolization, and elimination) and discrete events (drug intake). We use this toolkit to derive analytical expressions for blood concentration trajectories under various multi-dose regimens across several flagship pharmacokinetic models. We demonstrate that this mathematical framework furnishes an alternative and simplified way to model and retrieve analytical solutions for multi-dose dynamics. For instance, it enables the study of blood concentration responses to arbitrary dose regimens and facilitates the characterization of the long-term behavior of the solutions, such as their steady state.

Bioaccessibility of condensed tannins and their effect on the physico-chemical characteristics of lamb meat.

The bioaccessibility of tannins as antioxidants in meat is essential to maximise their effectiveness in protecting the product. This property determines the amount of tannins available to interact with meat components, inhibiting lipid and protein oxidation and, consequently, prolonging shelf life and preserving the sensory quality of the product. The objective of this study was to evaluate the bioaccessibility of condensed tannins (CT) from Acacia mearnsii extract (AME) and their effect on the physico-chemical characteristics of fattened lamb meat. Thirty-six Dorset × Hampshire lambs (3 months old and 20.8 ± 3.3 kg live weight) were used. The lambs were distributed equally (n = 9) into four treatments: T1, T2, T3 and T4, which included a basal diet plus 0%, 0.25%, 0.5% and 0.75% of CT from AME, respectively. At the end of the fattening period, bioaccessibility was evaluated, the animals were slaughtered and a sample of the longissimus dorsi (LD) muscle was collected to assess colour, lipid oxidation, cooking weight loss and shear force on days 1, 4, 7 and 14 of shelf-life, in samples preserved at -20 °C. In addition, the long chain fatty acid profile was analysed. A completely randomised design was used, and the means were compared with Tukey's test (P < 0.05). The mean lightness (L*), yellowness (b*) and hue (H*) values were higher for T3 and T4. The addition of CT did not affect (P > 0.05) redness (a*), cooking weight loss (CWL) or shear force (SF). T4 decreased (P < 0.05) stearic acid and increased cis-9 trans-12 conjugated linoleic acid (CLA). Bioaccessibility was higher in the supplemented groups (T1 < T2, T3 and T4). In conclusion, supplementing CT from AME in the diet of lambs did not reduce lipid oxidation, but T3 or T4 improved some aspects of meat colour and CLA deposition.

Recovery of Full Hyperextension After Anterior Cruciate Ligament Reconstruction Is Associated With Better Functional Outcomes but Greater Anterior Cruciate Ligament-Related Laxity After a Minimum 24-Month Follow-Up.

PURPOSE: To evaluate patients who had anterior cruciate ligament (ACL) reconstruction with preoperative hyperextension and compare physical examination (KT-1000 and pivot shift) and patient-reported outcome measures of patients who recovered the entire hyperextension with patients who did not. METHODS: Patients aged 18 to 60 years with more than 5° of knee hyperextension who had anatomic ACL reconstruction with any graft from June 2013 to June 2021 and at least a 24-month follow-up were evaluated retrospectively. Hamstrings and patellar tendon grafts were fixed around 20° to 30° of flexion and in full extension, respectively. Patients who could recover hyperextension were compared with patients who did not. Preoperative, intraoperative, and postoperative data, including physical examination and patient-reported outcome measures, were evaluated. RESULTS: Of the 225 patients evaluated, 48 (21.3%) did not recover hyperextension, and 177 recovered full range of motion. Patients who did not recover hyperextension had a larger graft diameter (8.7 ± 0.7 mm [confidence interval, 8.502-8.898 mm] vs 8.3 ± 0.7 mm [confidence interval, 8.197-8.403 mm]; P = .018). Regarding the postoperative subjective variables, patients who recovered hyperextension showed improvement on all scales (International Knee Documentation Committee, Lysholm, Forgotten Joint Score, and Global Perceived Effect) evaluated compared with patients who did not recover the range of motion. Patients who recovered hyperextension also showed more laxity on physical examination, measured by the KT-1000 (1.8 ± 0.8 vs 1.1 ± 1.0; P = .0006) and the pivot shift (62.1% [grade 0] and 37.9% [grade 1] vs 79.2% [grade 0] and 20.8% [grade 1]; P = .027). CONCLUSIONS: Patients with knee hyperextension who regained range of motion after ACL reconstruction have worse knee laxity than patients who regained full extension but not hyperextension. However, patients who recover full range of motion showed higher scores on subjective function scales, including a greater number of patients who achieved patient acceptable symptom state for the International Knee Documentation Committee score. LEVEL OF EVIDENCE: Level III, retrospective comparative therapeutic trial.

A Minimally Invasive Surgical Procedure to Harvest Palate Periosteum as a Source of Mesenchymal Stromal/Stem Cells for Bone Tissue Engineering.

The aim of this study is to validate a minimally invasive surgical procedure to harvest palate periosteum as a source of tissue for mesenchymal stromal/stem cells. We performed a standardized procedure to harvest the palate periosteum in ten subjects, which consisted of a 3 mm disposable punch and a Molt periosteal elevator to harvest a small full-thickness fragment of soft tissue at the hard palate area, between the upper bicuspids, 3 to 4 mm apical to the cement enamel junction. The one-third inner portion was fragmented, and following standard cell culture procedures, the adherent cells were cultured for three passages, after obtaining 70-90% confluence. Cell morphology analysis, flow cytometry analysis, and viability and osteogenic differentiation assays were performed. In all 10 cases, uneventful healing was observed, with no need for analgesic intake. The evaluation of cell morphology showed elongated spindle-shaped cells distributed in woven patterns. A high viability range was verified as well as an immunophenotype compatible with mesenchymal stem cell lineage. The differentiation assay showed the potential of the cells to differentiate into the osteogenic lineage. These results demonstrate that the minimally invasive proposed surgical technique is capable of supplying enough periosteum source tissue for stem cell culture and bone tissue engineering.

Stem cell-conditioned medium improves methylation patterns and quality of caprine preantral follicles.

This study investigated the methylation patterns of H3K4me3 and H3K9me3, as well as the mRNA expression of genes encoding the epigenetic regulators KDM1AX1, KDM1AX2, and KDM3A in goat preantral follicles developed in vivo (Uncultured control) or after in vitro culture for 7 days in either the absence (α-MEM+) or presence of conditioned medium (α-MEM+ + CM) from Wharton's jelly mesenchymal stem cells (WJ-MSCs). In the in vivo setting, all follicular categories exhibited similar H3K4me3 and H3K9me3 patterns, and transcripts of KDM1AX1, KDM1AX2, and KDM3A were detected in all samples. During in vitro culture, α-MEM+ + CM enhanced several important aspects. It increased the percentage of normal growing follicles, oocyte diameters across all categories, stromal cell density, and the H3K4me3 methylation pattern in preantral follicles. Simultaneously, it decreased the levels of reduced thiols and reactive oxygen species in the spent media, diminished the presence of lipofuscin aggresomes, lowered granulosa cell apoptotic rates, and reduced the H3K9me3 methylation pattern in preantral follicles. In conclusion, the findings from this study provide compelling evidence that supplementing the in vitro culture medium (α-MEM+) with CM from WJ-MSCs has a protective effect on goat preantral follicles. Notably, CM supplementation preserved follicular survival, as evidenced by enhanced follicular and oocyte growth and increased stromal cell density when compared to the standard culture conditions in the α-MEM+ medium. Furthermore, CM reduced oxidative stress and apoptosis and promoted alterations in H3K4me3 and H3K9me3 patterns.

Mesenchymal stem cells in animal reproduction: sources, uses and scenario.

Studies regarding mesenchymal stem cells turned up in the 1960's and this cell type created a great number of questions about its functions and applicability in science and medicine. When used with therapeutic intent, these cells present an inclination to migrate to sites of injury, inflammation or disease, where they secrete bioactive factors that stimulates the synthesis of new tissue. In this context, studies using rodents reported that MSCs promoted positive effects in the ovarian function in mice with premature aging of follicular reserve. In female bovines, experimental stem cell-based therapies have been used to either generate new oocytes with in vitro quality or stimulate such action in vivo. It is also reported, that the intraovarian application of mesenchymal stem cells generates a greater production of embryos in vitro and the production of early and expanded blastocysts. Additionally, analysis of ovarian tissue in animal subjected to treatment showed an increase in the number of developing follicles. Nevertheless, the treatments involving stem cells with different modes of application, different sources and different species were able to act on the hormonal, tissue, cellular and metabolic levels, generating positive results in the recovery and improvement of ovarian functions.

Estudos sobre células-tronco mesenquimais surgiram na década de 1960 e esse tipo de célula gerou muitas dúvidas sobre suas funções e aplicabilidade na ciência e na medicina. Quando utilizadas com intuito terapêutico, essas células apresentam tendência a migrar para locais de lesão, inflamação ou doença, onde secretam fatores bioativos que estimulam a síntese de novos tecidos. Nesse contexto, estudos utilizando roedores relataram que as CTM promoveram efeitos positivos na função ovariana em camundongos com envelhecimento precoce da reserva folicular. Em fêmeas bovinas, terapias experimentais baseadas em células-tronco têm sido utilizadas para gerar novos oócitos com qualidade in vitro ou estimular tal ação in vivo. É relatado também que a aplicação intraovariana de células-tronco mesenquimais gera maior produção de embriões in vitro e produção de blastocistos precoces e expandidos. Além disso, a análise do tecido ovariano em animais submetidos ao tratamento mostrou aumento no número de folículos em desenvolvimento. Apesar disso, os tratamentos envolvendo células-tronco com diferentes modos de aplicação, diferentes fontes e diferentes espécies foram capazes de atuar nos níveis hormonal, tecidual, celular e metabólico, gerando resultados positivos na recuperação e melhora das funções ovarianas.

A Survey of MPSoC Management toward Self-Awareness.

Managing Multi-Processor Systems-on-Chip (MPSoCs) is becoming increasingly complex as demands for advanced capabilities rise. This complexity is due to the involvement of more processing elements and resources, leading to a higher degree of heterogeneity throughout the system. Over time, management schemes have evolved from simple to autonomous systems with continuous control and monitoring of various parameters such as power distribution, thermal events, fault tolerance, and system security. Autonomous management integrates self-awareness into the system, making it aware of its environment, behavior, and objectives. Self-Aware Cyber-Physical Systems-on-Chip (SA-CPSoCs) have emerged as a concept to achieve highly autonomous management. Communication infrastructure is also vital to SoCs, and Software-Defined Networks-on-Chip (SDNoCs) can serve as a base structure for self-aware systems-on-chip. This paper presents a survey of the evolution of MPSoC management over the last two decades, categorizing research works according to their objectives and improvements. It also discusses the characteristics and properties of SA-CPSoCs and explains why SDNoCs are crucial for these systems.

Therapeutic dosage of isotretinoin in rats may influence orthodontic tooth movement.

Objective Isotretinoin, also known as 13-cis-retinoic acid, is an isomer of tretinoin, the oxidized form of Vitamin A. Orthodontic tooth movement (OTM) is the result of a cascade of inflammatory responses stimulated by a physical element that is the force generated by orthodontic appliances. Isotretinoin is mainly used among adolescents and young adults, and coincidentally it is this age group that also undergoes orthodontic treatment. Materials and Methods Fifty-five animals were used, and they were randomly divided into 11 groups, containing 5 animals in each group. Group 1: Control; Group 2: OTM for 7 days; Group 3: OTM for 14 days; Group 4: Treated with isotretinoin for 14 days with a dosage of 7.5 mg/kg/day; Group 5: Treated with isotretinoin for 14 days with a dosage of 1.0 mg/kg/day; Group 6: Treated with isotretinoin for 21 days with a dosage of 7.5 mg/kg/day; Group 7: Treated with isotretinoin for 21 days with a dosage of 1.0 mg/kg/day; Group 8: Treated with isotretinoin for 14 days with a dosage of 7.5 mg/kg/day and undergoing OTM for 7 days; Group 9: Treated with isotretinoin for 14 days with a dosage of 1.0 mg/kg/day and undergoing OTM for 7 days; Group 10: Treated with isotretinoin for 21 days with a dosage of 7.5 mg/kg/day and undergoing OTM for 14 days; Group 11: Treated with isotretinoin for 21 days with a dosage of 1.0 mg/kg/day and undergoing OTM for 14 days. In Groups 8, 9, 10 and 11, the animals were treated with isotretinoin for 7 days before OTM and maintained during the movement period in the respective groups. Results There was a significant difference in microtomographic parameters, including Trabecular Volume (BV/TV), Trabecular Thickness (Tb.Th), Number of Trabeculae (Tb.N), and Trabecular Separation (Tb.Sp), between the groups. The group that received orthodontic force in conjunction with isotretinoin treatment at a dosage of 7.5 mg/kg/day exhibited lower tooth displacement over a period of 21 days and 14 days. Conclusion Isotretinoin caused a reduction in tooth displacement during OTM when administered at a dose of 7.5 mg/kg/day and isotretinoin did change the microtomographic parameters of treated animals.

Optimal distribution grid allocation of reactive power with a focus on the particle swarm optimization technique and voltage stability.

A structured approach to managing reactive power is imperative within the context of power systems. Among the restructuring initiatives in the electrical sector, power systems have undergone delineation into three principal categories: generation, transmission, and distribution entities, each of which is overseen by an independent system operator. Notably, active power emerges as the predominant commodity transacted within the electrical market, with the autonomous grid operator assuming the responsibility of ensuring conducive conditions for the execution of energy contracts across the transmission infrastructure. Ancillary services, comprising essential frameworks for energy generation and delivery to end-users, encompass reactive power services pivotal in the regulation of bus voltage. Of particular significance among the array of ancillary services requisite in a competitive market milieu is the provision of adequate reactive power to uphold grid safety and voltage stability. A salient impediment to the realization of energy contracts lies in the inadequacy of reactive power within the grid, which poses potential risks to its operational safety and voltage equilibrium. The optimal allocation of the reactive power load is predicated upon presumptions of consistent outcomes within the active power market. Under this conceptual framework, generators are afforded continual compensation for the provision of reactive power indispensable for sustaining their active energy production endeavors.

Effect of poly-L-lactic acid and polydioxanone biostimulators on type I and III collagen biosynthesis.

OBJECTIVE: Safe, effective, and biocompatible minimally invasive procedures with the potential to stimulate collagen production have been made to recover dermal thickness and skin quality. The main of this animal model experiment was to observe the effect of poly-L-lactic acid (PLLA) and polydioxanone (PDO) biostimulators in collagen I and III after hypodermal injection. METHODOLOGY: Sixteen adult female rats (Wistar) were randomized into four groups and had dorsal treatment with: G1: hypodermic subcision (HS) only; G2: HS and PLLA hypodermic injection (HI), G3: HS and PDO HI; G4: Control, with no treatment. RESULTS: In histochemical, it was observed hypodermal and dermal tissue in more organized thickness in G3 and in G4 when compared to G1 and G2. There was few difference in G1 compared to G4. The tissue of G2 showed irregularities in the arrangement of collagen fibers, less defined structure and lower distribution of type I collagen compared to the other groups. There is a greater tendency for the proportions of type III collagen among tissues treated with both biostimulators (G2 and G3). PLLA and PDO had relatively similar percentages of collagen when compared to G4. The amount of type I collagen was higher in tissues treated with subcision, while type III collagen was higher in tissues treated with both biostimulators. CONCLUSION: G3 showed better performance in collagen production, although small, when compared with G2.

Role of Oxidants in Metal Extraction from Sulfide Minerals in a Deep Eutectic Solvent.

Metallurgical applications of deep eutectic solvents (DESs), known as ionometallurgy, have received significant research attention in recent years. While many studies claim that DESs are generally green and enhance process efficiency, others believe that industrial applications of ionometallurgy are generally not viable. Here, we report on leaching experiments of a sulfide flotation concentrate using ethaline, a chloride-based DES, in the presence of common oxidants. Following a mineral-based approach, we compare results with those obtained from aqueous chloride solutions to assess the influence of the leaching medium. We aim to contribute to a basic understanding of key differences between DESs and aqueous solutions and hope that this will help to make informed decisions about the suitability of DESs for leaching applications. Experiments were performed on a feed concentrate comprising a mixture of sulfide minerals along with substantial concentrations of Au, Ag, and Te. We found similar leaching behaviors for ethaline and aqueous solutions in nonoxidative leaching. However, when oxidizing agents were introduced, ethaline exhibited higher leaching efficiencies. Notably, the oxidation rate of pyrite in ethaline was very low, while chalcopyrite exhibited high oxidation rates. Furthermore, the results highlight significant variations in leaching rates depending on the type of oxidant, with the highest rate observed for I2, followed by CuCl2, and FeCl3. H2O2 and O2 were less effective. The leaching of gold-silver tellurides was possible in ethaline. This could be of particular significance, given that Au-Ag-Te compounds pose challenges in conventional cyanide treatment.

Combined approaches, including long-read sequencing, address the diagnostic challenge of HYDIN in primary ciliary dyskinesia.

Primary ciliary dyskinesia (PCD), a disorder of the motile cilia, is now recognised as an underdiagnosed cause of bronchiectasis. Accurate PCD diagnosis comprises clinical assessment, analysis of cilia and the identification of biallelic variants in one of 50 known PCD-related genes, including HYDIN. HYDIN-related PCD is underdiagnosed due to the presence of a pseudogene, HYDIN2, with 98% sequence homology to HYDIN. This presents a significant challenge for Short-Read Next Generation Sequencing (SR-NGS) and analysis, and many diagnostic PCD gene panels do not include HYDIN. We have used a combined approach of SR-NGS with bioinformatic masking of HYDIN2, and state-of-the-art long-read Nanopore sequencing (LR_NGS), together with analysis of respiratory cilia including transmission electron microscopy and immunofluorescence to address the underdiagnosis of HYDIN as a cause of PCD. Bioinformatic masking of HYDIN2 after SR-NGS facilitated the detection of biallelic HYDIN variants in 15 of 437 families, but compromised the detection of copy number variants. Supplementing testing with LR-NGS detected HYDIN deletions in 2 families, where SR-NGS had detected a single heterozygous HYDIN variant. LR-NGS was also able to confirm true homozygosity in 2 families when parental testing was not possible. Utilising a combined genomic diagnostic approach, biallelic HYDIN variants were detected in 17 families from 242 genetically confirmed PCD cases, comprising 7% of our PCD cohort. This represents the largest reported HYDIN cohort to date and highlights previous underdiagnosis of HYDIN-associated PCD. Moreover this provides further evidence for the utility of LR-NGS in diagnostic testing, particularly for regions of high genomic complexity.