Dung-beetles (Coleoptera, Scarabaeidae, Aphodiinae, Scarabaeinae) feeding on faeces of steppe marmots Marmotabobak (Rodentia, Sciuridae) in Middle Volga territory.

Background: In open terrestrial biomes of Holarctic realm, ground squirrels are recognised as keystone species inhabiting steppes. They shape the plant species composition and diversity and support a fauna of species associated with their burrows. Ground squirrels and associated dung-beetles are important elements of the steppe food webs, yet the trophic associations between species are still poorly studied. New information: The area in the northern outskirts of Obshchy Syrt plateau, on the border of Samara and Orenburg Provinces of Russia was surveyed and scarab beetles (Scarabaeidae) feeding on steppe marmot (Marmotabobak (Müller, 1776)) faeces were collected from six localities. Twenty eight species of two subfamilies - Aphodiinae and Scarabaeinae, - were identified with the majority of species belonging the genus Aphodius Hellwig, 1798. Seven species are recorded as consumers of marmot faeces for the first time. Only two nidicolous specialist species were found which suggests that the studied population of steppe marmots is as result of the recent secondary colonisation and not all the associated scarab beetle faunas were re-established.

Two Cases of Sporadic Amyotrophic Lateral Sclerosis With Contrasting Clinical Phenotypes: Genetic Insights.

Amyotrophic lateral sclerosis (ALS) is a fatal neuromuscular disease that affects individuals of diverse racial and ethnic backgrounds. There is currently no cure for ALS, and the number of efficient disease-modifying drugs for ALS is limited to a few, despite the large number of clinical trials conducted in recent years. The latter could be attributed to the significant heterogeneity of ALS clinical phenotypes even in their familial forms. To address this issue, we conducted postmortem genetic screening of two female patients with sporadic ALS (sALS) and contrasting clinical phenotypes. The results demonstrated that despite their contrasting clinical phenotypes, both patients had rare pathologic/deleterious mutations in five genes: ACSM5, BBS12, HLA-DQB1, MUC20, and OBSCN, with mutations in three of those genes being identical: BBS12, HLA-DQB1, and MUC20. Additional groups of mutated genes linked to ALS, other neurologic disorders, and ALS-related pathologies were also identified. These data are consistent with a hypothesis that an individual could be primed for ALS via mutations in a specific set of genes not directly linked to ALS. The disease could be initiated by a concerted action of several mutated genes linked to ALS and the disease's clinical phenotype will evolve further through accessory gene mutations associated with other neurological disorders and ALS-related pathologies.

A Rare Case of Penoscrotal Webbing and Extensive Hernias: An Anatomical Report With Genetic Insights.

During a routine anatomical dissection of an 81-year-old male cadaver received through the Gift Body Program of Saint Louis University School of Medicine (SLU SOM), a massive bulging in the abdominal area was observed that was consistent with numerous hernia repairs noted in the donor's self-reported medical history. Gross anatomical dissection of the cadaveric body revealed extensive herniation of portions of the small intestine and peritoneal sac along the costal margin and extending to the left aspect of the abdomen. Additionally, an uncircumcised phallus was buried within the suprapubic fat pad and demonstrated simple, grade III penoscrotal webbing (PSW), creating an impression of micropenis presence. To gain additional insights into the current case, analysis of the coding regions (exomes) of DNA procured from the body for putative genetic variants was performed using next-generation sequencing (NGS) technology. This analysis revealed 110 rare (minor allele frequency (MAF) ≤ 0.01), pathologic/deleterious genetic mutations. The most relevant variants to this case were the ones associated with male sexual development, BMP1 and BMP4; connective tissue development, COL3A1 and COL5A3; cilia morphogenesis and function, DNAH5 and MAPK15; as well as hormonal homeostasis, ESR1. Direct involvement of BMP1 both in male sexual development and hernia genesis makes it a strong candidate for linking the two pathologies, PSW and multiple hernias, observed in the present case. Yet the presence of a group of mutated genes linked to myopathies (ITGA7, NRAP, POLM, SCN5A, XIRP2) and muscular dystrophy (ITGA7) raises a question about the involvement of these muscular pathologies in hernia genesis and unsuccessful hernia repairs associated with the current case.

Amplicon metagenomics of dung beetles (Coleoptera, Scarabaeidae, Scarabaeinae) as a proxy for lemur (Primates, Lemuroidea) studies in Madagascar.

Dung beetles (Scarabaeidae, Scarabaeinae) are among the most cost-effective and informative biodiversity indicator groups, conveying rich information about the status of habitats and faunas of an area. Yet their use for monitoring the mammal species, that are the main providers of the food for the dung beetles, has only recently been recognized. In the present work, we studied the diet of four endemic Madagascan dung beetles (Helictopleurusfissicollis (Fairmaire), H.giganteus (Harold), Nanosagaboides (Boucomont), and Epilissussplendidus Fairmaire) using high-throughput sequencing and amplicon metagenomics. For all beetle species, the ⅔-¾ of reads belonged to humans, suggesting that human feces are the main source of food for the beetles in the examined areas. The second most abundant were the reads of the cattle (Bostaurus Linnaeus). We also found lower but significant number of reads of six lemur species belonging to three genera. Our sampling localities agree well with the known ranges of these lemur species. The amplicon metagenomics method proved a promising tool for the lemur inventories in Madagascar.

Combining structural and coevolution information to unveil allosteric sites.

Understanding allosteric regulation in biomolecules is of great interest to pharmaceutical research and computational methods emerged during the last decades to characterize allosteric coupling. However, the prediction of allosteric sites in a protein structure remains a challenging task. Here, we integrate local binding site information, coevolutionary information, and information on dynamic allostery into a structure-based three-parameter model to identify potentially hidden allosteric sites in ensembles of protein structures with orthosteric ligands. When tested on five allosteric proteins (LFA-1, p38-α, GR, MAT2A, and BCKDK), the model successfully ranked all known allosteric pockets in the top three positions. Finally, we identified a novel druggable site in MAT2A confirmed by X-ray crystallography and SPR and a hitherto unknown druggable allosteric site in BCKDK validated by biochemical and X-ray crystallography analyses. Our model can be applied in drug discovery to identify allosteric pockets.

Phylogenetic analysis of the Neotropical scarab beetle tribe Aegidiini (Coleoptera, Scarabaeidae, Orphninae) with description of new taxa.

In the Neotropics, orphnine scarab beetles are represented by the endemic tribe Aegidiini Paulian, 1984 with five genera and over 50 species. Phylogenetic analysis based on morphological characters of all supraspecific taxa of Orphninae showed that Aegidiini is comprised of two lineages. New subtribes, Aegidiina subtr. nov. (Aegidium Westwood, 1845, Paraegidium Vulcano et al., 1966, Aegidiellus Paulian, 1984, and Onorius Frolov & Vaz-de-Mello, 2015, and Aegidininasubtr. nov. (Aegidinus Arrow, 1904) are proposed to better reflect this phylogeny. Two new species of Aegidinus are described: A.alexanderisp. nov., from the Yungas in Peru and A.elbaesp. nov. from the Caqueta moist forests ecoregion in Colombia. A diagnostic key to Aegidinus species is given.

pIChemiSt ─ Free Tool for the Calculation of Isoelectric Points of Modified Peptides.

The isoelectric point (pI) is a fundamental physicochemical property of peptides and proteins. It is widely used to steer design away from low solubility and aggregation and guide peptide separation and purification. Experimental measurements of pI can be replaced by calculations knowing the ionizable groups of peptides and their corresponding pKa values. Different pKa sets are published in the literature for natural amino acids, however, they are insufficient to describe synthetically modified peptides, complex peptides of natural origin, and peptides conjugated with structures of other modalities. Noncanonical modifications (nCAAs) are ignored in the conventional sequence-based pI calculations, therefore producing large errors in their pI predictions. In this work, we describe a pI calculation method that uses the chemical structure as an input, automatically identifies ionizable groups of nCAAs and other fragments, and performs pKa predictions for them. The method is validated on a curated set of experimental measures on 29 modified and 119093 natural peptides, providing an improvement of R2 from 0.74 to 0.95 and 0.96 against the conventional sequence-based approach for modified peptides for the two studied pKa prediction tools, ACDlabs and pKaMatcher, correspondingly. The method is available in the form of an open source Python library at https://github.com/AstraZeneca/peptide-tools, which can be integrated into other proprietary and free software packages. We anticipate that the pI calculation tool may facilitate optimization and purification activities across various application domains of peptides, including the development of biopharmaceuticals.

Sex-Related Pain Behavioral Differences following Unilateral NGF Injections in a Rat Model of Low Back Pain.

Low back pain (LBP) is a globally prevalent and costly societal problem with multifactorial etiologies and incompletely understood pathophysiological mechanisms. To address such shortcomings regarding the role of neurotrophins in the underlying mechanisms of pain, an LBP model was developed in rats involving two unilateral intramuscular injections of nerve growth factor (NGF) into deep trunk muscles. To date, behavioral investigations of this NGF-LBP model have been limited, especially as it pertains to female pain behaviors. This study compared mechanical sensitivity to noxious (hyperalgesia) and non-noxious (hypersensitivity) stimuli in control and NGF-injected male and female rats through pain resolution. Although the baseline testing revealed no differences between males and females, NGF-injected females demonstrated prolonged ipsilateral deep trunk mechanical hyperalgesia that resolved seven days later than males. Moreover, females showed bilateral trunk mechanical sensitivity to noxious and non-noxious stimuli compared to only ipsilateral behaviors in males. Sex differences were also observed in the severity of behavioral responses, with females displaying greater mean differences from baseline at several timepoints. Overall, these NGF-LBP behavioral findings mirror some of the sex differences reported in the clinical presentation of LBP and accentuate the translatability of this NGF-LBP model. Future studies using this LBP-NGF model could help to elucidate the neurobiological mechanisms responsible for the development, severity, and/or resolution of muscular LBP as well as to provide insights into the processes governing the transition from acute to chronic LBP.

New taxa of Athyreacaridae (Acari: Heterostigmata) from Neotropical and Afrotropical realms.

A new genus and three new species of Athyreacaridae (Acari: Heterostigmata) are described associated with beetles of subfamily Bolboceratinae (Coleoptera: Geotrupidae) in Neotropical and Afrotropical realms: Neoathyreacarus pygmephoroides gen., sp. nov. collected on Bolborhinum tubericeps in Chile; Athyreacarus staturosus sp. nov. collected on Zefevazia cantisani  in Argentina; and Athyreacarus camerikae sp. nov. collected on Namibiobolbus helgae in South Africa. An updated key to genera and species of Athyreacaridae is provided. Athyreacarus staturosus sp. nov. is the largest ever described non-physogastric female among Heterostigmata.

Somatosensory behavioral alterations in a NGF-induced persistent low back pain model.

Low back pain (LBP) is a major global burden in part due to the underlying pathophysiological mechanisms being poorly understood. A LBP rat model involving two injections of nerve growth factor (NGF, an endogenous pain-related neurotrophin) into trunk musculature was recently developed. Additional behavioral work in this NGF-LBP rat model is required to better characterize local and remote somatosensory alterations related to NGF-induced peripheral and central sensitization. This work characterizes the time-dependent development of hypersensitivity to trunk and hindpaw cutaneous mechanical stimulation and deep muscle mechanical hyperalgesia in adult male Sprague-Dawley rats (n = 6/group). Behavioral assays were performed at baseline (Day 0, D0), D2, D5 (pre- and 4 h post-2nd NGF or control injection), D7, D10, and D14 in NGF and control groups. Trunk and hindpaw cutaneous mechanical hypersensitivity were tested using von Frey filaments. Deep trunk mechanical hyperalgesia was determined using a small animal algometer. NGF rats demonstrated increased cutaneous sensitivity to ipsilateral trunk mechanical stimuli at D7, D10, and D14. NGF rats also demonstrated ipsilateral deep mechanical hyperalgesia on D2, D5 + 4 h, D7, D10, and D14. Cutaneous hypersensitivity was delayed compared to deep hyperalgesia in NGF rats. No additional sensory changes were noted. Together, these results indicate that male mechanical somatosensory changes develop primarily locally in the ipsilateral trunk following unilateral NGF injections. These findings contrast with a previous report in female rats using this NGF-LBP model showing more widespread (bilateral) hyperalgesia and remote mechanical hypersensitivity. Future studies will examine potential sex-related pain behavioral differences in the NGF model.

Identification of larvae of two Aphodius Helwig, 1798 (Coleoptera: Scarabaeidae: Aphodiinae) species using morphology and DNA barcode.

The third instar larvae of Aphodius (Alocoderus) hydrochaeris (Fabricius, 1798) and A. (Bodilus) ictericus (Laicharting, 1781) are described based on scanning electron microscopy and COI sequences. COI barcode sequence for A. (A.) hydrohaeris is provided for the first time. Two haplotypes are discovered in A. (B.) ictericus.

Unraveling the Allosteric Cross-Talk between the Coactivator Peptide and the Ligand-Binding Site in the Glucocorticoid Receptor.

The glucocorticoid receptor (GR) is a nuclear receptor that controls critical biological processes by regulating the transcription of specific genes. There is a known allosteric cross-talk between the ligand and coregulator binding sites within the GR ligand-binding domain that is crucial for the control of the functional response. However, the molecular mechanisms underlying such an allosteric control remain elusive. Here, molecular dynamics (MD) simulations, bioinformatic analysis, and biophysical measurements are integrated to capture the structural and dynamic features of the allosteric cross-talk within the GR. We identified a network of evolutionarily conserved residues that enables the allosteric signal transduction, in agreement with experimental data. MD simulations clarify how such a network is dynamically interconnected and offer a mechanistic explanation of how different peptides affect the intensity of the allosteric signal. This study provides useful insights to elucidate the GR allosteric regulation, ultimately providing a foundation for designing novel drugs.

A new species of Aegidium Arrow (Coleoptera: Scarabaeidae: Orphninae) from Colombia.

In the Neotropical Region, Orphninae (Coleoptera: Scarabaeidae) are represented by the endemic tribe Aegidiini, which comprises five genera and 49 species (Paulian 1948; Colby 2009; Frolov Vaz-de-Mello 2015; Frolov et al. 2017a, 2017b, 2017c, 2019). Aegidium Westwood is the largest genus of the tribe and it comprises 24 valid species known from the southern Mexico in the north to Central Bolivia in the south (Frolov et al. 2015; Rojkoff Frolov 2017; Frolov Akhmetova 2020).

First record of Athyreacaridae (Acari: Heterostigmata) from Asia with description of two new species of Athyreacarus from India and Sri Lanka.

Mites of the family Athyreacaridae are recorded from Asia for the first time. Two new species of Athyreacarus are described, A. indicus sp. nov., collected on the beetles Bolbohamatum laevicolle and B. marginale in India, and A. brevisetosus sp. nov., collected on a beetle Bolboceras sp. in Sri Lanka. An updated key to species of the genus Athyreacarus is provided.

A new species of the endemic Madagascan scarab beetle genus, Madecorphnus grebennikovi (Coleoptera: Scarabaeidae: Orphninae): morphological description and DNA barcode.

A new species of the orphnine scarab beetle genus Madecorphnus Paulian, 1992, Madecorphnus grebennikovi Frolov, Akhmetova Vishnevskaya, new species, is described from the Marojejy National Park, Sava Region, northeastern Madagascar. The new species can be distinguished from the congeners by the parameres narrowly rounded in lateral view and having a small but distinct lateral teeth, and by the endophallic armature consisting of 1) a long straight sclerite with attached to its end a 2/3 shorted, somewhat curved sclerite, 2) separate smaller, elongate sclerite, and 3) a rather large area of microspinules. The 811 bp long fragment of the mitochondrial gene COI (DNA barcode) is provided as a part of the diagnosis of the new species. An updated key to the Madecorphnus species is given.

Description of a new brachypterous scarab species, Orphnus brevialatus (Coleoptera: Scarabaeidae: Orphninae) from East Africa, with notes on flightlessness in the orphnines.

A new brachypterous species of the scarab beetle genus Orphnus Macleay, 1819, O. (O.) brevialatus Frolov Akhmetova, new species, is described from Kipengere Range (Livingstone Mountains), Tanzania. The new species differs from other flightless Orphnus species occurring in East Africa in having a well-developed pronotal armature with two long, acute lateral processes. In the shape of the head, pronotum, and male genitalia it is similar to O. compactilis Quedenfeldt, 1882, but differs from it, in addition to the reduced wings and shorter body, in the shape of the endophallic armature. Flightlessness is rather common among the orphnines: 56 orphnine species out of 235 have the wings reduced to different degree making them incapable of flight, although this is a preliminary minimum figure. Flightless species are found in Hybalus Dejean, 1833, Chaetonyx Schaum, 1862, Orphnus Macleay, Hybaloides Quedenfeldt, 1884, Aegidium Westwood, 1845 and Stenosternus Karsch, 1881. All regional orphnine faunas except for Indo-Malayan and Madagascan include flightless species, although flightlessness is more common in African taxa.

The taxonomic status of Aphodius insperatus Petrovitz, A. longipennis Rakovic and A. inylchekensis Frolov (Coleoptera: Scarabaeidae: Aphodiinae).

The subgenus Bodilus Mulsant Rey, 1870 of the genus Aphodius Helwig, 1798 sensu lato comprises 30 species in the Palearctic fauna (Frolov 2001, 2002, Dellacasa et al. 2016 [treated as genus Bodilus]). Frolov (2001) reviewed the species of this subgenus from Russia and adjacent countries and suggested that the monotypic subgenus Paramelinopterus Rakovic, 1984 shares the main diagnostic characters of Bodilus. Frolov (2001) was unable, however, to examine the type species of Paramelinopterus, A. longipennis Rakovic, 1984, at that time. Král (2016) suggested that A. (Bodilus) inylchekensis Frolov, 2001 is a junior synonym of A. longipennis. Recently we have re-examined all the types as well as the additional material accumulated in the museums and can confirm that A. inylchekensis is a distinct species, but A. insperatus Petrovitz, 1967 and A. longipennis cannot be reliably separated by the morphological characters. In the present contribution we illustrate the major diagnostic characters of the species based on the type specimens and also provide a distribution map of the known localities of the species.

Adapting free energy perturbation simulations for large macrocyclic ligands: how to dissect contributions from direct binding and free ligand flexibility.

Large and flexible ligands gain increasing interest in the development of bioactive agents. They challenge the applicability of computational ligand optimization strategies originally developed for small molecules. Free energy perturbation (FEP) is often used for predicting binding affinities of small molecule ligands, however, its use for more complex ligands remains limited. Herein, we report the structure-based design of peptide macrocycles targeting the protein binding site of human adaptor protein 14-3-3. We observe a surprisingly strong dependency of binding affinities on relatively small variations in substituent size. FEP was performed to rationalize observed trends. To account for insufficient convergence of FEP, restrained calculations were performed and complemented with extensive REST MD simulations of the free ligands. These calculations revealed that changes in affinity originate both from altered direct interactions and conformational changes of the free ligand. In addition, MD simulations provided the basis to rationalize unexpected trends in ligand lipophilicity. We also verified the anticipated interaction site and binding mode for one of the high affinity ligands by X-ray crystallography. The introduced fully-atomistic simulation protocol can be used to rationalize the development of structurally complex ligands which will support future ligand maturation efforts.

Sagittal Craniosynostosis with Uncommon Anatomical Pathologies in a 56-Year-Old Male Cadaver.

Sagittal craniosynostosis (CS) is a pathologic condition that results in premature fusion of the sagittal suture, restricting the transverse growth of the skull leading in some cases to elevated intracranial pressure and neurodevelopmental delay. There is still much to be learned about the etiology of CS. Here, we report a case of 56-year-old male cadaver that we describe as sagittal CS with torus palatinus being an additional anomaly. The craniotomy was unsuccessful (cephalic index, CI = 56) and resulted in abnormal vertical outgrowth of the craniotomized bone strip. The histological analysis of the latter revealed atypical, noncompensatory massive bone overproduction. Exome sequencing of DNA extracted from the cadaveric tissue specimen performed on the Next Generation Sequencing (NGS) platform yielded 81 genetic variants identified as pathologic. Nine of those variants could be directly linked to CS with five of them targeting RhoA GTPase signaling, with a potential to make it sustained in nature. The latter could trigger upregulated calvarial osteogenesis leading to premature suture fusion, skull bone thickening, and craniotomized bone strip outgrowth observed in the present case.

A review of the Malagasy species of the genus Trigonochilus Brenske (Coleoptera: Scarabaeidae: Rutelinae: Adoretini) with description of a new species.

Africa and Madagascar have a large and diverse fauna of the chafers of the ruteline beetle tribe Adoretini (Ohaus 1912, 1918). Many nominal taxa from Madagascar are still poorly known and many new ones were described recently (Akhmetova Montreuil 2010; Montreuil 2010, 2013; Montreuil Frolov 2014; Frolov Montreuil 2018). One of the poorly known Adoretini taxa is the genus Trigonochilus Brenske, 1896. Trigonochilus was proposed to accommodate the East-African species T. coriaceus Brenske, 1896, based on the enlarged and curved metatibiae and a short, spur-like process of the labrum (Brenske 1896). Arrow (1901) noted that the enlarged metatibiae of this species (described from a single male) is a secondary sexual character and added another species to this genus, T. politus Arrow, 1901, from Angola. These species were revised by Krikken (1979). Fairmaire (1903) described Adoretus oedipus from Madagascar (without any precise locality) and noted its enlarged metatibia and the shape of the labrum similar to that in Trigonostonum Burmeister, 1844. Fairmaire was apparently unaware of the Brenske's and Arrow's works as he did not discuss the possible affinities of A. oedipus with Trigonochilus species. Ohaus (1912) moved A. oedipus to Trigonochilus after examination of the type series but he did not discuss the characters of the species in detail as well as its distribution on the island. Therefore, the genus Trigonochilus Brenske comprised three species described so far, two from Africa and one from Madagascar.