Genetic and phenotypic attributes of splenic marginal zone lymphoma.

Splenic marginal zone B-cell lymphoma (SMZL) is a heterogeneous clinico-biological entity. The clinical course is variable, multiple genes are mutated with no unifying mechanism, and essential regulatory pathways and surrounding microenvironments are diverse. We sought to clarify the heterogeneity of SMZL by resolving different subgroups and their underlying genomic abnormalities, pathway signatures, and microenvironment compositions to uncover biomarkers and therapeutic vulnerabilities. We studied 303 SMZL spleen samples collected through the IELSG46 multicenter international study (NCT02945319) by using a multiplatform approach. We carried out genetic and phenotypic analyses, defined self-organized signatures, validated the findings in independent primary tumor metadata and in genetically modified mouse models, and determined correlations with outcome data. We identified 2 prominent genetic clusters in SMZL, termed NNK (58% of cases, harboring NF-κB, NOTCH, and KLF2 modules) and DMT (32% of cases, with DNA-damage response, MAPK, and TLR modules). Genetic aberrations in multiple genes as well as cytogenetic and immunogenetic features distinguished NNK- from DMT-SMZLs. These genetic clusters not only have distinct underpinning biology, as judged by differences in gene-expression signatures, but also different outcomes, with inferior survival in NNK-SMZLs. Digital cytometry and in situ profiling segregated 2 basic types of SMZL immune microenvironments termed immune-suppressive SMZL (50% of cases, associated with inflammatory cells and immune checkpoint activation) and immune-silent SMZL (50% of cases, associated with an immune-excluded phenotype) with distinct mutational and clinical connotations. In summary, we propose a nosology of SMZL that can implement its classification and also aid in the development of rationally targeted treatments.

Gross anatomical description of the extrinsic and intrinsic scapular and brachial muscles of Sapajus apella (Linnaeus, 1758).

BACKGROUND: The robust brown capuchin monkey (Sapajus apella) is a South American primate with preferences for arboreal locomotion, which requires specific thoracic limb muscle adaptations. The present investigation studied the gross anatomy of the extrinsic and intrinsic scapular and brachial muscles. METHODS: Gross dissections were performed in both thoracic limbs of four formaldehyde-fixed specimens. RESULTS: Three rhomboideus muscles were present (capitis, cervicis, and thoracis). The trapezius muscle was divided into two parts (cervicis and thoracis). The pectoralis abdominalis and omotransversarius muscles were present. The anconeus muscle was found as an individual muscle or fused to the caput mediale of the triceps brachii muscle. The brachialis muscle had among one and two heads. The anconeus epitrochlearis was absent. CONCLUSION: These muscles of Sapajus apella are adapted for arboreal locomotion and some terrestrial habits, since these have many similarities with other primates with a similar locomotor patterns.

Evaluation of the MD Anderson tumor score for diffuse large B-cell lymphoma in the rituximab era.

OBJECTIVES: Diffuse large B-cell lymphoma (DLBCL) is an aggressive heterogeneous lymphoma with standard treatment. However, 30%-40% of patients still fail, so we should know which patients are candidates for alternative therapies. IPI is the main prognostic score but, in the rituximab era, it cannot identify a very high-risk (HR) subset. The MD Anderson Cancer Center reported a score in the prerituximab era exclusively considering tumor-related variables: Tumor Score (TS). We aim to validate TS in the rituximab era and to analyze its current potential role. METHODS: From GELTAMO DLBCL registry, we selected those patients homogeneously treated with R-CHOP (n = 1327). RESULTS: Five-years PFS and OS were 62% and 74%. All variables retained an independent prognostic role in the revised TS (R-TS), identifying four different risk groups, with 5-years PFS of 86%, 71%, 50%, and very HR (28%). With a further categorization of three variables of the original TS (Ann Arbor Stage, LDH and B2M), we generated a new index that allowed an improvement in HR assessment. CONCLUSIONS: (a) All variables of the original TS retain an independent prognostic role, and R-TS remains predictive in the rituximab era; (b) R-TS and additional categorization of LDH, B2M, and AA stage (enhanced TS) increased the ability to identify HR subsets.

Gross anatomy of the shoulder and arm intrinsic muscles in the white-footed tamarin (Saguinus leucopus - Günther, 1876): Inter- and intraspecific anatomical variations.

BACKGROUND: Saguinus leucopus is a Neotropical primate with an arboreal quadrupedal locomotion pattern, which requires wide movements of the shoulder and arm. This investigation studies the muscles of these regions in order to serve as a basis for clinical and surgical procedures and to compare with other primates. METHODS: Gross dissections of twenty thoracic limbs were performed. RESULTS: The muscles examined were the deltoid, supraspinatus, infraspinatus, subscapularis, teres major, teres minor, coracobrachialis longus, coracobrachialis brevis, biceps brachii, brachialis, triceps brachii, tensor fasciae antebrachii, and anconeus epitrochlearis. The anconeus was absent. The following variants were found: an accessory head of the biceps brachii, the unilateral absence of the short head of the biceps brachii, an accessory head of the coracobrachialis longus, and one infraspinatus muscle innervated by the axillary nerve. CONCLUSIONS: These muscles are adapted to quadrupedal locomotion and can have inter- and intraspecific variations in their attachments and innervation.

Morphometric, anatomic and radiographic study of the scapula in the white-footed tamarin (Saguinus leucopus): report of scapular cartilage and one variation in cranial (superior) transverse scapular ligament.

The white-footed tamarin (Saguinus leucopus) is an endangered endemic primate of Colombia, mainly due to the deforestation of its habitat and illegal trade, which generates a high incidence of these animals in wildlife care centres. Musculoskeletal system disorders in S. leucopus are one of the most common diseases and therefore the aim of this study was to contribute to the morphologic studies with a morphometric, anatomic and radiographic description of the scapula in this species to provide a basis for medical interventions, surgical approaches, radiologic diagnoses and comparative functions of this bone. Gross dissections of each scapular region were made in eight specimens without a diagnosis of osteomuscular disease. These specimens died from natural cases in the wildlife care centres of the Corporación Autónoma Regional de Caldas (CORPORCALDAS); after necropsy their carcasses were fixed with 10% formaldehyde, 5% mineral oil and 1% phenic acid in these centres over the course of at least 1 week. X-rays of the scapula were taken in the small animal clinic of the Universidad del Tolima, and morphometric data of the scapulae were obtained with a digital calliper. The scapula of the white-footed tamarin was a flat triangular bone with a deep scapular notch in its cranial margin, where there was a cranial transverse scapular ligament that was absent in two specimens. The coracoid process was highly developed, medially covering the humeral joint. The dorsal margin was covered by the scapular cartilage, which was highly developed in the caudal angle. In the dorsal fourth of the caudal margin, there was a surface from which the m. teres major originated. The lateral surface had a scapular spine with a long hamatus process of the acromion until the lateral part of the humeral joint. The infraspinatus fossa was wider than the supraspinous fossa. On the costal surface, the subscapular fossa was formed by three subscapular lines and one subscapular ridge, the latter helping to form the surface for the m. teres major. In the two radiographic views, caudocranial to the scapula and dorsoventral to the thorax, the scapular spine, acromion, coracoid process, scapular incisura, supraglenoid tubercle, caudal margin, subscapular ridge, and the joints with the clavicle and the humerus could be observed. The scapula of the white-footed tamarin presented bony reliefs that share characteristics with other primates but also with domestic mammals due to its quadrupedal locomotion, which allowed us to correlate its morphologic adaptation with its quadrupedal arboreal displacement.

Actinobacillus seminis GroEL-homologous protein agglutinates sheep erythrocytes.

Actinobacillus seminis, a commensal of ovine and caprine reproductive organs, is able to induce epididymitis in the small ruminants that it infects. In this work, we characterised two protein bands of approximately 150 kDa and 65 kDa. These proteins cross-reacted with a polyclonal serum against Gallibacterium anatis hemagglutinin and with a polyclonal serum from sheep with epididymitis, indicating that the proteins are expressed in vivo; the two proteins also interacted with biotin-labeled sheep fibrinogen and fibronectin, suggesting that they may function as adhesins. The participation of these proteins as adhesins was confirmed by a cultured human bladder cell-A. seminis adhesion assay and adherence inhibition by preincubation of A. seminis with polyclonal antiserum to the 150 kDa protein. Both proteins presented sequence identity with an A. seminis GroEL protein by mass spectrometry analysis and agglutinated glutaraldehyde-fixed sheep red blood cells. Immunogold labeling was observed by transmission electron microscopy on bacterial cells that were negatively stained, and a peroxidase reaction was detected in A. seminis biofilms, when an anti-A. seminis 150 kDa protein serum was used, indicating the presence of this protein on the surface of A. seminis and in biofilms. The A. seminis GroEL-homologue is a multifunctional protein that likely acts as a hemagglutinin.

Anatomic description of the palmaris longus muscle and report of variant nerve supply in the white-footed tamarin (Saguinus leucopus Günther, 1876).

The gross anatomy of the palmaris longus muscle in 12 specimens of Saguinus leucopus was studied by dissection. This muscle was constant and unipennate, originated in the medial epicondyle of the humerus; inserted onto flexor retinaculum and palmar aponeurosis; and innerved by the median nerve, and in one case, together with the ulnar nerve. This is the first report of double innervation for this muscle in primates.

Impact on prognosis of the regional distribution of MGMT methylation with respect to the CpG island methylator phenotype and age in glioma patients.

Clinical and molecular prognostic factors in gliomas include age, IDH mutation, the glioma CpG island methylator phenotype (G-CIMP+) and promoter methylation of the O(6)-methylguanine DNA-methyltransferase (MGMT) gene. Among these markers, a predictive value was reported in glioblastomas (GBM) for MGMT promoter methylation, in particular in elderly GBM patients. In this study, methylation data from 46 glioma samples with the Illumina 450K platform were obtained and extended using external data to include a total of 247 glioma samples. Methylation analysis of the whole MGMT gene with this platform revealed two strongly survival-associated CpG regions within the promoter and the gene body, which were confirmed in a reported dataset of high grade-gliomas. Methylation at the promoter (CpG 25, cg12981137 and the prognostic model MGMT-STP27) and at the gene body CpG 165 (cg07933035), were significantly associated with better overall survival, and strongly correlated with G-CIMP+ status. In this series, the prognostic value of MGMT methylation at the promoter was not observed in G-CIMP- cases, although around 50 % of them were MGMT-methylated. These results were also obtained in an homogeneously-treated series of chemoradiated G-CIMP- GBMs analyzed by MSP and qMSP, and confirmed in a reported pyrosequencing-analyzed series of gliomas. Interestingly, in contrast to the MGMT promoter, gene body methylation was of prognostic value in G-CIMP-patients older than 65 years. Our study highlights the relevance of the prognostic value of the different regions of methylation throughout the MGMT gene that could be affected by specific G-CIMP profiles and age groups.

Expression of COX-2 on Reed-Sternberg cells is an independent unfavorable prognostic factor in Hodgkin lymphoma treated with ABVD.

Cyclooxygenase 2 (COX-2) is an inflammatory enzyme involved in the pathogenesis and prognosis of several malignancies. In the present study, we investigated the prognostic value of COX-2 expression in a large (N = 242), uniformly treated Hodgkin lymphoma (HL) population from the Spanish Network of HL using tissue microarrays. Univariate and multivariate analysis was done, including comparing the most recognized clinical variables: the early- and advanced-stage subgroups. COX-2 was expressed on Reed-Sternberg cells in 37% of patients. There were no differences in the distribution of clinical variables according to COX-2 expression. With a median follow-up time of 58 months, PFS at 5 years was 60% and 79% for COX-2(+) and COX-2(-) patients, respectively (P = .003). The overall survival was 73% and 91%, respectively (P < .001). The major impact on prognosis was observed in the early AA stage (I-II) group. In fact, in these low-risk groups the expression of COX-2 defined a group with significantly worse progression-free and overall survival. In conclusion, COX-2 was expressed on Reed-Sternberg cells in one-third of HL patients and was a major independent, unfavorable prognostic factor in early-stage HL. We conclude that COX-2 may be a major prognostic variable in HL and a potential therapeutic target.

Anomalous origin of the right subclavian artery in a Neotropical otter (Lontra longicaudis, Olfers-1818).

The Neotropical otter (Lontra longicaudis) is a mustelid distributed geographically from Mexico to Argentina. Anomalous origins of the aortic arch branches are rarely reported in wild carnivorans. Therefore, this study aimed to report the anomalous branching of the aortic arch in one formaldehyde-fixed specimen of L. longicaudis. The aortic arch provided three branches: the bicarotid trunk and the left and right subclavian arteries. The latter passed dorsally to the esophagus toward the right side without a mark of compression at the esophagus. This is the first report of an anomalous origin of the right subclavian artery in L. longicaudis.

Barriers and Facilitators to Implementing a Community-Based Psychosocial Support Intervention Conducted In-Person and Remotely: A Qualitative Study in Quibdó, Colombia.

Community-based psychosocial support group (CB-PSS) interventions using task-shifting approaches are well suited to provide culturally appropriate services in low- and middle-income countries. However, contextual barriers and facilitators must be considered to tailor interventions effectively, particularly considering the challenges introduced by the COVID-19 pandemic. We explore the barriers, facilitators, and psychosocial changes associated with implementing a CB-PSS group intervention delivered by local lay providers to conflict-affected adults in Quibdó, Colombia, using both in-person and remote modalities. Data were analyzed from 25 individual interviews with participants and a focus group discussion involving staff members, including 7 community psychosocial agent facilitators and 2 mental health professional supervisors. The analysis used a thematic approach grounded in a descriptive phenomenology to explore the lived experiences of participants and staff members during implementation. Participant attendance in the in-person modality was compromised by factors such as competing work and family responsibilities and disruption caused by the COVID-19 pandemic. Participants in the remote modality faced challenges concerning unstable Internet connectivity, recurrent power outages caused by heavy rain, distractions, interruptions, and threats to confidentiality by family and coworkers. Despite these challenges, data revealed key contextual facilitators, including the community-based knowledge of facilitators and integration of traditional practices, such as the comadreo (informal talks and gatherings). Respondents shared that the CB-PSS groups promoted stronger community relationships and created opportunities for participants to exchange peer support, practice leadership skills, develop problem-solving skills based on peers' experiences, and enhance emotional regulation skills. Differences and similarities across in-person and remote modalities are discussed, as are key considerations for practitioners and policymakers.

Evolutionary anatomical and functional characteristics of the intrinsic shoulder and brachial muscles in the northern anteater (Tamandua mexicana).

Tamandua mexicana is an anteater species native from Mexico to Peru. This species is of great evolutionary interest because it belongs to one of the oldest clades of placental mammals in the American continent. This study aimed to describe the origin, insertion, and arterial supply of the intrinsic shoulder and brachial muscles of T. mexicana. We also compared the masses of the functional groups. Gross dissections were performed on both thoracic limbs of 13 cadavers. ANOVA followed by Tukey's test was used for statistical analyses. The subscapularis muscle presents a hiatus to the common tendon of the caput breve of the biceps brachii and coracobrachialis muscles. A variant accessory muscle, the m. articularis humeri lateralis, was found on the lateral surface of the shoulder joint. M. deltoideus pars acromialis has two bellies. The teres major muscle is perforated by the aponeurotic origin of the m. tensor fasciae antebrachii. The triceps brachii has two capita longi. The caput mediale is fused with the m. anconeus medialis. The caput laterale can have an accessory belly as an anatomical variant. Among the functional groups, a significant difference was found between the elbow extensors and flexors, with the latter having the lowest mass. In conclusion, the intrinsic muscles of T. mexicana presented unique features for the species, as well as arrangements in mass distribution that evidence a possible evolutionary convergence among species of the Superorder Xenarthra.

A genetic profiling guideline to support diagnosis and clinical management of lymphomas.

The new lymphoma classifications (International Consensus Classification of Mature Lymphoid Neoplasms, and 5th World Health Organization Classification of Lymphoid Neoplasms) include genetics as an integral part of lymphoma diagnosis, allowing better lymphoma subclassification, patient risk stratification, and prediction of treatment response. Lymphomas are characterized by very few recurrent and disease-specific mutations, and most entities have a heterogenous genetic landscape with a long tail of recurrently mutated genes. Most of these occur at low frequencies, reflecting the clinical heterogeneity of lymphomas. Multiple studies have identified genetic markers that improve diagnostics and prognostication, and next-generation sequencing is becoming an essential tool in the clinical laboratory. This review provides a "next-generation sequencing" guide for lymphomas. It discusses the genetic alterations of the most frequent mature lymphoma entities with diagnostic, prognostic, and predictive potential and proposes targeted sequencing panels to detect mutations and copy-number alterations for B- and NK/T-cell lymphomas.

Evaluation of the Effectiveness of Geogrids Manufactured from Recycled Plastics for Slope Stabilization-A Case Study.

This study aimed to investigate the sustainable use of recycled plastics, specifically polypropylene (PP) and high-density polyethylene (HDPE), in the manufacture of geogrids for geotechnical and civil engineering applications. Plastics were collected from a recycling center, specifically targeting containers used for food, cleaning products, and other domestic packaging items. These plastics were sorted according to the Möbius triangle classification system, with HDPE (#2) and PP (#5) being the primary categories of interest. The research methodologically evaluates the mechanical properties of PP/HDPE (0/100, 25/75, 50/50, 75/25 and 100/0% w/w) composites through tensile and flexural tests, exploring various compositions and configurations of geogrids. The results highlight the superiority of pure recycled HDPE processed into 1.3 mm thick laminated yarns and hot air welded for 20 to 30 s, exhibiting a deformation exceeding 60% in comparison to the PP/HDPE composites. Through SolidWorks® Simulation, it was shown that the adoption of a trigonal geogrid geometry optimizes force distribution and tensile strength, significantly improving slope stabilization efficiency. Based on the results obtained, a laboratory-scale prototype geogrid was developed using an extrusion process. The results underscore the importance of careful composite design and yarn configuration selection to achieve the desired mechanical properties and performance in geogrid applications. It emphasizes the potential of recycled plastics as a viable and environmentally friendly solution for stabilizing slopes, contributing to the reduction in plastic waste and promoting sustainable construction practices.

miRNA expression in diffuse large B-cell lymphoma treated with chemoimmunotherapy.

Diffuse large B-cell lymphoma (DLBCL) prognostication requires additional biologic markers. miRNAs may constitute markers for cancer diagnosis, outcome, or therapy response. In the present study, we analyzed the miRNA expression profile in a retrospective multicenter series of 258 DLBCL patients uniformly treated with chemoimmunotherapy. Findings were correlated with overall survival (OS) and progression-free survival (PFS). miRNA and gene-expression profiles were studied using microarrays in an initial set of 36 cases. A selection of miRNAs associated with either DLBCL molecular subtypes (GCB/ABC) or clinical outcome were studied by multiplex RT-PCR in a test group of 240 cases with available formalin-fixed, paraffin-embedded (FFPE) diagnostic samples. The samples were divided into a training set (123 patients) and used to derive miRNA-based and combined (with IPI score) Cox regression models in an independent validation series (117 patients). Our model based on miRNA expression predicts OS and PFS and improves upon the predictions based on clinical variables. Combined models with IPI score identified a high-risk group of patients with a 2-year OS and a PFS probability of < 50%. In summary, a precise miRNA signature is associated with poor clinical outcome in chemoimmunotherapy-treated DLBCL patients. This information improves upon IPI-based predictions and identifies a subgroup of candidate patients for alternative therapeutic regimens.

Codeletion of 1p and 19q determines distinct gene methylation and expression profiles in IDH-mutated oligodendroglial tumors.

Oligodendroglial tumors (OTs) are primary brain tumors that show variable clinical and biological behavior. The 1p/19q codeletion is frequent in these tumors, indicating a better prognosis and/or treatment response. Recently, the prognostically favorable CpG island methylator phenotype (CIMP) in gliomas (G-CIMP+) was associated with mutations in the isocitrate dehydrogenase 1 and isocitrate dehydrogenase 2 (IDH) genes, as opposed to G-CIMP- tumors, highlighting the relevance of epigenetic mechanisms. We performed a whole-genome methylation study in 46 OTs, and a gene expression study of 25 tumors, correlating the methylation and transcriptomic profiles with molecular and clinical variables. Here, we identified two different epigenetic patterns within the previously described main G-CIMP+ profile. Both IDH mutation-associated methylation profiles featured one group of OTs with 1p/19q loss (CD-CIMP+), most of which were pure oligodendrogliomas, and a second group with intact 1p/19q and frequent TP53 mutation (CIMP+), most of which exhibited a mixed histopathology. A third group of OTs lacking the CIMP profile (CIMP-), and with a wild-type IDH and an intact 1p/19q, similar to the G-CIMP- subgroup, was also observed. The three CIMP groups presented a significantly better (CD-CIMP+), intermediate (CIMP+) or worse (CIMP-) prognosis. Furthermore, transcriptomic analyses revealed CIMP-specific gene expression signatures, indicating the impact of genetic status (IDH mutation, 1p/19q codeletion, TP53 mutation) on gene expression, and pointing to candidate biomarkers. Therefore, the CIMP profiles contributed to the identification of subgroups of OTs characterized by different prognoses, histopathologies, molecular features and gene expression signatures, which may help in the classification of OTs.

Origin and Distribution of the Brachial Plexus in Two Procyonids (Procyon cancrivorus and Nasua nasua, Carnivora).

Procyon cancrivorus and Nasua nasua are two procyonids with different evolutionary adaptations to use their thoracic limbs. Therefore, this study aimed to characterize the differences in the brachial plexus between both species. Five P. cancrivorus and five N. nasua cadavers were used to perform this investigation with the permission of the bioethics committee and environmental license. Gross dissections were performed on the cervical, pectoral, and thoracic limb regions to find the origin and distribution of the brachial plexus. The brachial plexus of both species originated in a variant manner from C5-T1, C5-T2, C6-T1, or C6-T2. All brachial plexus nerves were observed and, interestingly, the musculocutaneous sent a communicating branch to the median nerve medially to the axillary artery, forming an ansa axillaris in both species. An ansa pectoralis was also observed medially to the axillary artery. Additionally, in P. cancrivorus, the musculocutaneous nerve innervates the pronator teres and flexor carpi radialis muscles and communicates with the median nerve at the elbow level to continue as a common trunk at the antebrachium. The brachial plexus has differences between both procyonids, although in both species, it could conserve a primitive arrangement present within the infraorder Arctoidea.

Detection of pyrimidine-rich DNA sequences based on the formation of parallel and antiparallel triplex DNA and fluorescent silver nanoclusters.

In this work, the use of DNA-stabilized fluorescent silver nanoclusters for the detection of target pyrimidine-rich DNA sequences by formation of parallel and antiparallel triplex structures is studied by molecular fluorescence spectroscopy. In the case of parallel triplexes, the probe DNA fragments are Watson-Crick stabilized hairpins, and whereas in the case of antiparallel triplexes, the probe fragments are reverse-Hoogsteen clamps. In all cases, the formation of the triplex structures has been assessed by means of polyacrylamide gel electrophoresis, circular dichroism, and molecular fluorescence spectroscopies, as well as multivariate data analysis methods. The results have shown that it is possible the detection of pyrimidine-rich sequences with an acceptable selectivity by using the approach based on the formation of antiparallel triplex structures.

Evolutionary comparative analysis of the extrinsic thoracic limb muscles in three procyonids (Procyon cancrivorus Cuvier, 1798, Nasua nasua Linnaeus, 1766, and Potos flavus Schreber, 1774) based on their attachments and innervation.

The procyonids (Procyon cancrivorus, Nasua nasua and Potos flavus) are Neotropical carnivorans with the ability to climb trees; however, each one has different locomotor preferences. Thereby, P. flavus is highly arboreal, P. cancrivorus is mainly terrestrial with abilities to swim, and N. nasua is also fossorial. These activities not only require movements of the hands but stabilize the thoracic limb, an action performed by the extrinsic muscles. Besides, former descriptions performed in procyonid species have obsolete terms for these muscles, generating confusion about the comparison among species. Thereby, muscle innervation has also been used to support the evolutionary derivation of the muscles. Therefore, this study aimed to describe the attachments and innervations of these muscles in three procyonids. There were intra- and interspecific anatomical variations in the attachments of all extrinsic thoracic limb muscles. However, based on the innervation, several evolutionary derivations in procyonids could be found, such as: the cleidobrachialis muscle derived from the deltoideus muscle; the atlantoscapularis muscle of P. flavus derived from the serratus ventralis cervicis muscle; the pectoralis transversus muscle derived from the pectoralis profundus and superficiales muscles; and the pectoralis abdominalis muscle derived from the cutaneus trunci muscle. Some functions could be associated with locomotor habits, among them a highly developed pectoralis abdominalis in Nasua for its fossorial habits and the atlantoscapularis in Potos for its arboreal and prehensile habits. Thus, the extrinsic muscles in procyonids have evolved for locomotor preferences, but mainly due to their phylogenetic relationship within the family Procyonidae.