Anna E. Romanovich

The systematic placement of the enigmatic monotypic geometrid genus Cheimoptena Danilevsky, 1969 has long been contentious. We used a multi-locus approach, analysing eight molecular markers (COI, GAPDH, RpS5, wingless, EF-1α, ArgK, Ca-ATPase, and H3) to clarify its phylogenetic position. Preliminary analysis revealed an error in the GenBank database: a COI barcode identical to Cheimoptena pennigera was found in the published mitogenome of the saturniid Neoris haraldi, suggesting the species’ presence in northwestern China. Subsequent reconstructions with comprehensive taxonomic sampling consistently placed Cheimoptena within a strongly supported clade comprising the tribes Campaeini, Alsophilini, and Prosopolophini. Although Cheimoptena is firmly nested within this complex, it is difficult to assign it to a specific tribe due to topological instability across trees. We addressed the potential impact of long-branch attraction, concluding that the clustering of Cheimoptena with this tribal complex is robustly driven by a genuine phylogenetic signal rather than methodological artefacts, while internal branching remains sensitive to outgroup selection. In terms of morphology, Cheimoptena exhibits a mosaic of characters, sharing similarities in genital structures with Alsophilini and pupal features with Prosopolophini. We confirm Cheimoptena represents an isolated, early-diverging lineage within this complex and maintain its status as a distinct tribe, Cheimoptenini.

Objective: Type A cystinuria is caused by mutations in the SLC3A1 gene encoding rBAT, the heavy subunit of the rBAT-b^0,+AT transporter. Exon 8 of SLC3A1 gene encodes a region of the rBAT ectodomain involved in hydrophobic-core formation and stabilization of its subdomain architecture. Missense mutations in this region can disrupt its maturation in endoplasmic reticulum (ER) and promote ER-associated degradation (ERAD). The aim of this study was to characterize the structural consequences of frequent exon 8 missense variants, evaluate their effects on rBAT-b^0,+AT maturation, and assess their potential as targets for pharmacological chaperone therapy. Methods: Four amino acid substitutions in rBAT ectodomain (p.Met467Thr, p.Met467Lys, p.Ser455Leu and p.Arg456Cys) were analysed. rBAT missense variant interpretation was based on ClinVar, gnomAD, and published functional studies. Structural modeling was performed using AlphaFold 3, with visualization in UCSF Chimera and contact analysis in YASARA. Results and Discussion: Three of the four variants (M467T, M467K, and R456C) were predicted to significantly destabilize the rBAT structure, whereas S455L showed only a modest effect on global stability. The most pronounced effects were observed for M467T and M467K, which disrupted packing of the hydrophobic core. S455L mainly affected surface packing through increased hydrophobicity and loss of potential hydrogen bonds. R456C introduced a free cysteine residue that may promote aberrant disulfide bond formation. Conclusions: The identified variants are predicted to destabilize the rBAT ectodomain while likely preserving early rBAT-b^0,+AT assembly. Impaired ectodomain maturation may trigger ER quality-control mechanisms and ERAD of the complex. These variants therefore represent potential targets for future pharmacological chaperone-based approaches in type A cystinuria.

Цистинурия — наследственное заболевание, характеризующееся нарушением транспорта цистина и развитием рецидивирующего нефролитиаза. Представлен клинический и молекулярно-генетический анализ случая цистинурии у молодой женщины. Описание клинического случая: пациентка 24 лет, страдающая псориазом и мочекаменной болезнью с дебютом заболевания в возрасте 18 лет. В 2023 г., в возрасте 21 года, у пациентки были выявлены правосторонний гидронефроз и множественные конкременты чашечно-лоханочной системы правой почки. Было проведено этапное урологическое лечение: стентирование правого мочеточника в апреле 2023 г. и перкутанная нефролитолапаксия с лазерной литотрипсией в октябре 2023 г. В январе 2024 г., в возрасте 22 лет, было выполнено повторное перкутанное вмешательство на правой почке с достижением «stone-free» статуса. Анализ состава удаленных конкрементов показал содержание цистина 95–100%. Экскреция цистина с мочой варьировала от 267,98 до 1197,4 мг/сут. Молекулярно-генетическое исследование (панель генов нефролитиаза) выявило в гене SLC7A9 гетерозиготный патогенный вариант c.614dupA (p.Asn206GlufsTer3) и новый, ранее не описанный миссенс-вариант c.1328A>G (p.Ser379Gly), классифицированный как вариант с неопределенным клиническим значением (VUS).Диагноз цистинурии типа B подтвержден совокупностью клинических, биохимических и молекулярно-генетических данных. Выявление известного патогенного варианта и нового ранее не описанного варианта c.1328A>G (p.Ser379Gly), рассматриваемого как потенциально патогенный, но в настоящее время классифицированного как VUS, подчеркивает значение молекулярно-генетической диагностики для верификации цистинурии и необходимость дальнейших исследований редких генетических вариантов. Представленный случай также демонстрирует важность индивидуального междисциплинарного подхода к ведению пациентов с сопутствующей аутоиммунной патологией (псориазом).

Cystinuria is an inherited disorder caused by pathogenic variants in SLC3A1 or SLC7A9, leading to impaired tubular reabsorption of cystine and recurrent cystine stone formation. Disease severity is determined by genotype, the degree of cystinuria, and patient adherence to long-term preventive therapy. Cases with long-term follow-up and molecular genetic confirmation are of particular clinical interest. We report a 36-year-old woman with severe recurrent cystine nephrolithiasis who was followed for 12 years after disease onset at the age of 24 years, shortly after her second pregnancy. Despite standard preventive measures, including high fluid intake, sodium and animal protein restriction, urinary alkalinization, and repeated attempts at standard-dose D-penicillamine therapy, the disease remained highly refractory. During follow-up, the patient experienced 6 surgical interventions and 10 episodes of spontaneous stone passage. Stone analysis repeatedly confirmed 100% cystine composition. Urinary cystine excretion ranged from 169.8 to 979 mg/day. Sanger sequencing identified a pathogenic homozygous SLC3A1 variant, c.1400TC (p.Met467Thr), associated with severe impairment of rBAT subunit trafficking and protein folding defects. Following poor tolerance of conventional thiol therapy doses, continuous low-dose D-penicillamine (250 mg/day) was initiated in February 2024 and was well tolerated. With this regimen, combined with dietary measures and high fluid intake, no new stones were detected on CT imaging over nearly 2 years of follow-up. This case illustrates a severe genotype-associated cystinuria phenotype linked to homozygous SLC3A1 c.1400TC (p.Met467Thr) and suggests that long-term low-dose D-penicillamine may be an effective and better tolerated therapeutic option in patients who cannot tolerate standard dosing regimens.

Pieris mannii (Mayer, 1851) has been repeatedly reported in the literature for countries in the Caucasus region (southern Russia, Georgia, Armenia, and Azerbaijan). However, most, if not all, of these records are the result of misidentifications due to significant morphological similarity of P. mannii with P. napi pseudorapae Verity, 1908, a southwest Asian subspecies of the Green-veined White P. napi (Linnaeus, 1758). This problem does not exist for butterflies from Western Europe, as the nominotypical subspecies of Pieris napi native to Western Europe is clearly distinguished from P. mannii by its wing pattern. In this study, based on morphological and DNA barcoding analysis of multiple Pieris specimens, we provide the first reliable evidence of the presence of P. mannii in Georgia. We also demonstrate that, despite significant intraspecific differentiation, DNA barcoding will allow for the unambiguous identification of P. mannii across different parts of its range, including the Caucasian region.

The genus Morpheis Hübner, [1820] (Lepidoptera: Cossidae) represents a taxonomically challenging group within Zeuzerinae, characterised by uniform, simply structured genitalia and significant intraspecific variability in external morphology, specifically in wing pattern, body colouration, and size. The taxonomic status and phylogenetic placement of many Morpheis taxa remain uncertain. Our study provides the first comprehensive taxonomic revision of Morpheis, based on detailed morphological analysis, examination of type specimens, assessment of distribution records, and molecular data. As a result, here we provide the first complete taxonomic list with updated diagnoses and distribution maps, based on 1247 specimen records and 191 publicly available observations. We describe the female genitalia of Morpheis for the first time and provide illustrations of imagoes for all currently recognised species, as well as of available type specimens. Additionally, we describe the immature stages and summarise the trophic associations of known Morpheis species. Based on morphological and molecular data, we recognise 11 valid Morpheis species. We applied molecular-based species delimitation methods, which uncovered far more candidate species within the genus than are recognised by traditional taxonomy, suggesting overlooked cryptic diversity that morphology alone cannot detect. We conclude that the integrative approach used here provides a powerful tool, especially valuable for understudied Zeuzerinae groups with high intraspecific variability in traditional characters.

BACKGROUND: The CRISPR/Cas9 technology enables the generation of genetically modified founder animals (F0 generation) already at the stage of zygote editing. However, the resulting offspring are often mosaic, meaning they carry different mutations in different cells, which complicates accurate genotyping using standard tissue samples. To establish a stable knockout line, it is necessary to identify F0 individuals carrying the target mutations, which requires subsequent crossing with wild-type mice and a large-scale analysis of the F1 offspring, involving significant costs. Therefore, the efficient prediction of inheritable mutations at the F0 generation stage is a critical task. AIM: A comparative evaluation of the efficiency of the TIDE and ICE bioinformatic algorithms for analyzing Sanger sequencing data to accurately predict inheritable mutations in the vldlr gene in F0 mosaic mice. METHODS: The study was conducted on five F0 mosaic mice with mutations in the vldlr gene, obtained by microinjection of CRISPR/Cas9 components into zygotes. Genomic DNA was isolated from ear tissue, the target region of the vldlr gene was amplified by PCR and sequenced by the Sanger method. The resulting chromatograms were analyzed using the TIDE and ICE algorithms. The predictions were validated by crossing the F0 mice with wild-type mice and analyzing the inheritance of mutations in the F1 generation. RESULTS: The comparison of the algorithms showed that both programs correctly predicted all mutations that were subsequently detected in the F1 generation. In total, 31 F1 offspring were analyzed. CONCLUSION: Both tools are suitable for primary screening. The analysis of the offspring confirmed that all actually inherited mutations were predicted by both methods.

Several lineages within the Eriophyoidea, an ancient group of phytoparasitic acariform mites, produce large protective silk webs on plant surfaces. This study examined the diversity and phylogeny of web-spinning eriophyoids associated with the leaves of Mangifera indica (mango) in Asia, Africa, and North America and Lepisanthes rubiginosa (mertajam) in Vietnam. We report, for the first time, silk-producing structures in Aceria aegyptindicae and Aculops knorri, as well as the presence of tibial seta l’ I and a lobular unpaired prosomal gland in Cisaberoptus kenyae. Molecular and morphological data indicate female dimorphism in A. knorri and reveal cryptic diversity within Cisaberoptus and mango-associated Aceria. The web-spinning species C. kenyae and Ac. aegyptindicae form a strongly supported clade and frequently coexist in syntopy on mango leaves, likely co-producing silk web. Phylogenetic analyses of COI and 28S genes place this clade as sister to Cecidophyinae, while grouping Aberoptus and A. knorri with Anthocoptini and Aceriini. These results demonstrate convergence, cryptic speciation, and unexpected mutualism in web-spinning eriophyoids, providing a framework for future research on the evolution of silk producing organs in this economically important mite group.

The psyllid genus Cacopsylla comprises mainly bisexually reproducing species; however, some members of this genus exhibit a unisexual mode of reproduction. Using an integrative approach that combines molecular and cytogenetic methods, as well as Wolbachia screening, we conducted a comprehensive study of the Palaearctic species C. ledi. We show that this species uses various reproductive strategies (bisexual and parthenogenetic) across its distribution range. Our findings indicate that the bisexual mode of reproduction has emerged at least twice in the evolutionary history of C. ledi. Bisexual populations in southern Fennoscandia are of ancestral origin, whereas the bisexual mode of reproduction observed in northern Fennoscandia represents a recent secondary transition from parthenogenesis. We report that in the first case, parthenogenetic and bisexual lineages can be easily distinguished not only cytogenetically but also by DNA barcoding, while in the second case, “bisexual” individuals share DNA barcodes with parthenogenetic ones. A comprehensive Wolbachia screening (1140 specimens across the entire distribution range) revealed Wolbachia infection in every specimen of C. ledi, indicating a significant role of the endosymbiont in the biology and evolution of this species.