D2 Region Research Articles

Genetic and environmental impact on variation in the palatal dimensions in permanent dentition: a twin study.

The objective of this study was to assess the relative contributions of genetic and environmental factors to variation in palatal parameters in twins with completed maxillary growth. The subjects of this study comprised digital dental casts of 50 monozygotic and 35 dizygotic twin pairs. The subjects' average age was 17.95 ± 2.83years. Zygosity determination was carried out using 15 specific DNA markers and an amel fragment of the amelogenin gene. The interdental distances were measured between selected dental landmarks at the occlusal and gingival planes. The palatal height, surface area and volume were measured between the gingival plane and the midpalate suture. High heritability estimates were observed for all transverse intra-arch measurements. The palate height (a2 = 0.8), dental arch width in the molar area (a2 = 0.86), palatal surface area (a2 = 0.61) and palate volume (a2 = 0.69) were under strong additive genetic control. Moderate genetic dominance was observed for dental arch widths at the gingival line in the canine (d2 = 0.5) and premolar regions (d2 = 0.78-0.81). Sexual dimorphism was shown, with males exhibiting a greater arch width, palate surface area and volume than females (p < 0.01). The majority of palate parameters variation in twins was controlled by genetic effects, and most were highly heritable.

Distribution, morphological and molecular characterization of root-lesion nematodes (Pratylenchus spp.) (Tylenchida: Pratylenchidae) in maize (Zea mays L.) (Poales: Poaceae) in Türkiye.

Root-lesion nematodes (RLN) are the most economically important pathogenic nematodes attacking maize. Significant economic losses due to lesion nematodes have been reported in maize producing countries in the world. This study was conducted to determine the distribution and identity of root-lesion nematodes (Pratylenchus spp.) (Tylenchida: Hoplolaimidae) in maize (Zea mays L.) (Poales: Poaceae) fields of the Black Sea region of Türkiye. For this purpose, 39 locations were surveyed and soil samples were taken from 17 regional provinces. Nematodes were extracted using the modified Baerman funnel technique. The species were identified based on sequences of the Internal Transcribed Spacer (ITS) region of ribosomal DNA, as well as morphological characters and morphometrics. In addition, species identifications were confirmed using species-specific primers in the D3 expansion region of 26S rDNA. At the end of the study, 51.3% of the maize production areas sampled in the region were infected with root-lesion nematode species. Pratylenchus agilis, P. mediterraneus, P. neglectus, P. penetrans, P. thornei, and P. vulnus were identified, and were present in 25%, 5%, 25%, 10%, 15%, and 20% of samples, respectively. To our knowledge, this is the first report of P. agilis in Türkiye. The present study concluded that the molecular analysis of Pratylenchus sequences based on the ITS and D3 region of ribosomal RNA genes allowed the identification of six root lesion nematode species. This study is of great importance in terms of adding additional species to the root-lesion nematode fauna in Turkey and will provide data for future research on the management of these nematodes.

Penghapusan Pembantu Pegawai Pencatat Nikah (P3N) Terhadap Pelayanan Pernikahan

This study examines the impact of removing P3N on marriage services at the Padang Sidempuan city Office of Religious Affairs after the fall of the instruction of the Directorate General of Islamic Community Guidance number Dj.II/113 of 2009 regarding the use PNBP funds for marital reconciliation and P3N management, as well as the instruction of the Director General of Islamic Community Guidance number Dj.II/1 0f 2015 about the P3N designation for regions D1 and D2. The research method used is field research with a qualitative approach.The primary data from the instruction of the Directorate General of Islamic Community Guidance number Dj.II/113 of 2009, instruction of the Director General of Islamic Community Guidance number Dj.II/1 0f 2015, Head of KUA Padangsidimpuan Selatan district, Padangsidimpuan Tenggara district, the Padangsidimpuan Angkola Julu district, P3N who was dismissed and the community. And the secondary data in this study are books, journals, theses and other supporting sources.Data collection techniques with observation, interviews, and documentation, with qualitative descriftive data analysis techniques. As for the reserach results obtained in this study P3N removal is carried out in two stages, namely 1) all P3N in the working area of KUA Padangsidimpuan city were officially abolished since the fall of intruction of the Director General of Islamic Community Guidance number Dj.II/113 of 2009 and 2) was not reappointed after an instruction from the Director General of Islamic Community Guidance number Dj.II/I of 2015 because the entire area of KUA Padangsidimpuan city is classified as typology C. 1) The impact of removing P3N is that it has an impact on KUA which has one who doubles as the head of KUA. 2)There was a time conflict between someone who wanted to consult with the time of registration of marriage outside the KUA. 3) Information that is slow to reach the public. 4) Wedding schedule shift. 5)P3N is still used by the public.

Fungi associated with the Asian citrus psyllid, Diaphorina citri Kuwayama, in northeastern Mexico

ABSTRACT Diaphorina citri Kuwayama (Asian citrus psyllid, ACP) is a devastating pest due to transmission of Liberibacter spp. to citrus plants. We identified presumed entomopathogenic fungi (EF) naturally associated with field-collected adult ACP, both killed (mycosed) and alive, at 14 field sites in the Gulf Coastal Plain of NE Mexico. On dead ACP, five species of EF (Hypocreales) were observed: Hirsutella citriformis Speare (Ophiocordycipitaceae), Cordyceps javanica (Frieder. & Bally) Kepler, Shrestha & Spatafora, Beauveria cf. bassiana (Bals.) Vuill., and Akanthomyces cf. lecanii (Cordycipitaceae); also, Niveomyces J.P.M. Araújo & C. de Bekker (Cordycipitaceae), a hyperparasite of Ophiocordycipitaceae, in this case H. citriformis. The most prevalent fungi on field ACP were H. citriformis and C. javanica, on dead and on live, incubated insects, respectively. In laboratory bioassays, C. javanica and H. citriformis conidia from cultures killed >60% of ACP adults. H. citriformis, C. javanica, and A. lecanii killed >60% nymphs. Niveomyces was observed only growing on H. citriformis fructifications on field-killed ACP; it was absent from live incubated insects, indicating Niveomyces is a mycoparasite on H. citriformis. Niveomyces cultures (ARSEF 10230) on agar grew as a Pleurodesmospora synanamorph; its LSU rRNA region D1–D2 sequences (Genbank OM780101) were 99.81% identical to Niveomyces sequences. Conidia from Niveomyces cultures were entomopathogenic on ACP nymphs and adults. These are the first reports of Niveomyces associated with H. citriformis; of the in vitro dimorphic growth (synanamorphs) Niveomyces/Pleurodesmospora; and of experimental verification of entomopathogenicity of Niveomyces. Understanding the interactions of these fungi will support sustainable ACP management.

Correlation between clinical phenotypes and genotypes among 46 children with SCN1A-related developmental epileptic encephalopathy

To explore the correlation between clinical phenotypes and genotypes among 46 children with SCN1A-related developmental epileptic encephalopathy (DEE). Clinical data of 46 children with DEE and SCN1A variants identified at the Guangzhou Women and Children's Medical Center between January 2018 and June 2022 were collected. The children were grouped based on their age of onset, clinical manifestations, neurodevelopmental status, and results of genetic testing. The correlation between SCN1A genotypes and clinical phenotypes was analyzed. Among the 46 patients, 2 children (4.35%) had developed the symptoms before 3 months of age, 42 (91.30%) were between 3 to 9 months, and 2 cases (4.35%) were after 10 months. Two cases (4.35%) presented with epilepsy of infancy with migrating focal seizures (EIMFS), while 44 (95.7%) had presented with Dravet syndrome (DS), including 28 cases (63.6%) with focal onset (DS-F), 13 cases (29.5%) with myoclonic type (DS-M), 1 case (2.27%) with generalized type (DS-G), and 2 cases (4.55%) with status epilepticus type (DS-SE). Both of the two EIMFS children had severe developmental delay, and among the DS patients, 7 cases had normal development, while the remaining had developmental delay. A total of 44 variants were identified through genetic sequencing, which included 16 missense variants and 28 truncating variants. All EIMFS children had carried the c.677C>T (p.Thr226Met) missense variant. In the DS group, there was a significant difference in the age of onset between the missense variants group and the truncating variants group (P < 0.05). Missense variants were more common in D1 (7/15, 46.7%) and pore regions (8/15, 53.3%), while truncating variants were more common in D1 (12/28, 42.9%). Children with variants outside the pore region were more likely to develop myoclonic seizures. The clinical phenotypes of DEE are diverse. There is a difference in the age of onset between individuals with truncating and missense variants in the SCN1A gene. Missense variants outside the pore region are associated with a higher incidence of myoclonic seizures.

Phylogenetic study about Korean Asobara Foerster (Hymenoptera: Braconidae: Alysiinae) species, with description of a new species

A new species in the genus Asobara Foester, (Braconidae: Alysiinae), Asobara rotundata sp. nov., is described and illustrated. In addition, we used the barcode region of mitochondrial cytochrome c oxidase I (COI), cytochrome c oxidase II (COII), cytochrome c (Cyt b), NADH dehydrogenase subunit 1 (ND1), and D2 region of the 28S nuclear gene (28S rDNA) to analyze the phylogeny of the available Korean Asobara species and identify a new species. These molecular analyses indicated that A. rotundata sp. nov. was recovered independently from the other Asobara species. Our results support that Asobara japonica occupies the most basal position of this genus and is a sister clade of other Asobara species. This also indicated that A. leveri is a sister clade to the A. rossica clade. An identification key for the Asobara species recorded in Korea is provided.

Anaerobic gut fungal communities in marsupial hosts.

The anaerobic gut fungi (AGF) inhabit the alimentary tracts of herbivores. In contrast to placental mammals, information regarding the identity, diversity, and community structure of AGF in marsupials is extremely sparse. Here, we characterized AGF communities in 61 fecal samples from 10 marsupial species belonging to four families in the order Diprotodontia: Vombatidae (wombats), Phascolarctidae (koalas), Phalangeridae (possums), and Macropodidae (kangaroos, wallabies, and pademelons). An amplicon-based diversity survey using the D2 region of the large ribosomal subunit as a phylogenetic marker indicated that marsupial AGF communities were dominated by eight genera commonly encountered in placental herbivores (Neocallimastix, Caecomyces, Cyllamyces, Anaeromyces, Orpinomyces, Piromyces, Pecoramyces, and Khoyollomyces). Community structure analysis revealed a high level of stochasticity, and ordination approaches did not reveal a significant role for the animal host, gut type, dietary preferences, or lifestyle in structuring marsupial AGF communities. Marsupial foregut and hindgut communities displayed diversity and community structure patterns comparable to AGF communities typically encountered in placental foregut hosts while exhibiting a higher level of diversity and a distinct community structure compared to placental hindgut communities. Quantification of AGF load using quantitative PCR indicated a significantly smaller load in marsupial hosts compared to their placental counterparts. Isolation efforts were only successful from a single red kangaroo fecal sample and yielded a Khoyollomyces ramosus isolate closely related to strains previously isolated from placental hosts. Our results suggest that AGF communities in marsupials are in low abundance and show little signs of selection based on ecological and evolutionary factors.IMPORTANCEThe AGF are integral part of the microbiome of herbivores. They play a crucial role in breaking down plant biomass in hindgut and foregut fermenters. The majority of research has been conducted on the AGF community in placental mammalian hosts. However, it is important to note that many marsupial mammals are also herbivores and employ a hindgut or foregut fermentation strategy for breaking down plant biomass. So far, very little is known regarding the AGF diversity and community structure in marsupial mammals. To fill this knowledge gap, we conducted an amplicon-based diversity survey targeting AGF in 61 fecal samples from 10 marsupial species. We hypothesize that, given the distinct evolutionary history and alimentary tract architecture, novel and unique AGF communities would be encountered in marsupials. Our results indicate that marsupial AGF communities are highly stochastic, present in relatively low loads, and display community structure patterns comparable to AGF communities typically encountered in placental foregut hosts. Our results indicate that marsupial hosts harbor AGF communities; however, in contrast to the strong pattern of phylosymbiosis typically observed between AGF and placental herbivores, the identity and gut architecture appear to play a minor role in structuring AGF communities in marsupials.

Aliinostoc bakau sp. nov. (Cyanobacteria, Nostocaceae), a New Microcystin Producer from Mangroves in Malaysia

A new microcystin-producing mangrove cyanobacterium, Aliinostoc bakau sp. nov., was isolated from a tropical mangrove in Penang, Malaysia, and characterized using combined morphological and phylogenetic approaches. Cultures were established in liquid media of different salinities (0, 7, 14, 21, 28, and 35 ppt). Optimal growth observed at both 7 and 14 ppt was consistent with the origin of the strain from an estuarine mangrove environment. Phylogenetic analysis based on the 16S rRNA gene strongly indicated that the strain is a member of the genus Aliinostoc and is distinct from other currently sequenced species in the genus. The sequences and secondary structure of the 16S–23S ITS region D1–D1’ and Box–B helices provided further confirmation that the new species is clearly distinct from previously described Aliinostoc species. Amplification of the mcyE gene fragment associated with the production of microcystin in A. bakau revealed that it is identical to that in other known microcystin-producing cyanobacteria. Analysis of the extracts obtained from this strain by HPLC-MS/MS confirmed the presence of microcystin variants (MC-LR and -YR) at concentrations of 0.60 μg/L and MC-RR at a concentration of 0.30 μg/L. This is the first record of microcystin production from Aliinostoc species in tropical mangrove habitats.

Diversity and toxicity of the planktonic diatom genus Pseudo-nitzschia from coastal and offshore waters of the Southeast Pacific, including Pseudo-nitzschia dampieri sp. nov.

To expand knowledge of Pseudo-nitzschia species in the Southeast Pacific, we isolated specimens from coastal waters of central Chile (36°S–30°S), the Gulf of Corcovado, and the oceanic Robinson Crusoe Island (700 km offshore) and grew them into monoclonal strains. A total of 123 Pseudo-nitzschia strains were identified to 11 species based on sequencing of the ITS region of the nuclear rDNA and on ultrastructural and morphometric analyses of the frustule in selected representatives of each clade: P. australis, P. bucculenta, P. cf. chiniana, P. cf. decipiens, P. fraudulenta, P. hasleana, P. multistriata, P. plurisecta, P. cf. sabit, the new species P. dampieri sp. nov., and one undescribed species. Partial 18S and 28S rDNA sequences, including the hypervariable V4 and D1–D3 regions used for barcoding, were gathered from representative strains of each species to facilitate future metabarcoding studies. Results showed different levels of genetic, and at times ultrastructural, diversity among the above-mentioned entities, suggesting morphological variants (P. bucculenta), rapidly radiating complexes with ill-defined species boundaries (P. cf. decipiens and P. cf. sabit), and the presence of new species (P. dampieri sp. nov., Pseudo-nitzschia sp. 1, and probably P. cf. chiniana). Domoic acid (DA) was detected in 18 out of 82 strains tested, including those of P. australis, P. plurisecta, and P. multistriata. Toxicity varied among species mostly corresponding to expectations from previous reports, with the prominent exception of P. fraudulenta; DA was not detected in any of its 10 strains tested. In conclusion, a high diversity of Pseudo-nitzschia exists in Chilean waters, particularly offshore.

Classification and identification of mosquitoes in China based on rDNA 28S D5 region

Accurate classification and identification of mosquitoes are essential for the prevention and control of mosquito-borne diseases. In this study, adult mosquitoes were collected from 15 cities across 14 provinces in China. They were identified morphologically with the dominant species determined. Furthermore, representative samples were identified at the molecular level based on rDNA 28S D5. In total, 880 adult mosquitoes were collected belonging to Culex (266), Aedes (473), Armigeres (13), and Anopheles (5). Aedes albopictus and “C. pipiens subgroup” were the dominant species. A total of 140 sequences of 28S D5 region (68 for “C. pipiens subgroup”, 51 for Ae. albopictus, 18 for Ar. subalbatus, and three for An. sinensis) ranging from 148 to 161 bp were obtained, with 100 % success of amplification and sequencing. Molecular identification were consistent with morphological classification. Sequence analysis showed that “C. pipiens subgroup” was identified into three clades: the traditional C. pipiens subgroup (Clade I), the newly discovered C. cf. perexiguus (Clade II), and C. new sp. (Clade III). Clade I contained the most abundant haplotypes (16) widely distributed without geographical differences. Clade II included six haplotypes that were aggregately distributed south of the Yangtze River. Only three sequences in Clade III showed two haplotypes with no geographical differences. Further morphological comparisons demonstrated differences in body color, beaks, and abdomens among the three clades. In conclusion, the rDNA 28S D5 region could effectively distinguish Culex, Aedes, Armigeres, and Anopheles species at the lower category level, demonstrating its potential as a mini-DNA barcode for mosquito identification.

MOLECULAR IDENTIFICATION AND ANTIBACTERIAL ACTIVITY OF MACROFUNGUS Trametes sanguineus (L.)

Aim. The molecular identification of Pycnoporus sanguineus, a previously morphologically mushroom, was done to see the antibacterial activity against pathogenic bacteria Staphylococcus aureus and Salmonella typhi. Methods. A fragment of the D2 region of 28S rDNA was amplified by PCR, sequenced, and BLAST was performed using the consensus sequence. The maximum identity score was used to build a phylogenetic tree. Agar well diffusion was used to study the antibacterial activity. Results. Sequencing of a 700 base pair PCR amplicon was carried and a 616 base pair of D2 region of large subunit gene was generated. The 100 blast hits on the D2 region of LSU gene showed similarity to Trametes sanguineus voucher PRSC95 (GenBank Accession Number: JN164795.1) based on nucleotide homology and phylogenetic analysis. Antibacterial screening revealed that the crude extract had higher activity on Staphylococcus aureus, with a 3mm to 13mm zone of inhibition and a 100µg minimum inhibitory concentration, compared to Salmonella typhi. Salmonella typhi had a 5 mm to 15 mm zone of inhibition and a 200 µg minimum inhibitory concentration. Conclusion. According to the obtained result, the morphologically identified mushroom Pycnoporus sanguies can be referred to as Trametes sanguine, and it can be used for producinig antibacterial agents.

Phylogenetic Position of the Genus Manunema (Nematoda, Plectida, Leptolaimidae)

During an investigation of a natural sandy beach on Jeju Island, South Korea, a species of marine nematode belonging to the rare genus Manunema was discovered and is now being reported. Up until now, no molecular sequences of this genus have been available, and its position within the nematode system has been a subject of dispute. Currently, Manunema is classified within the order Plectida, either as the only genus of the family Peresianidae in the superfamily Haliplectoidea or directly as a genus of the family Leptolaimidae. Here, we report on the discovery of Manunema cf. proboscidis Gerlach, 1957, which was found off Jeju Island, South Korea. We also conduct an in-depth investigation of its peculiar morphology, with a particular focus on the reproductive system type observed within the species. Additionally, we examine the phylogenetic relationship of the genus using 18S rRNA sequences. Partial mtCOI, nearly complete sequences of the 18S ribosomal RNA gene as well as a partial 28S (D2–D3 region) gene are provided. Morphologically, the presence of tubular precloacal supplements (shared by superfamily Leptolaimoidea and not Haliplectoidea) and antidromously reflected ovaries of females in Manunema (shared by the order Plectida and not Desmoscolecida) may have taxonomic significance. Our phylogenetic trees based on 18S rRNA also reflected this, including Manunema in a well-supported monophyletic clade of the family Leptolaimidae.

Preparation and Application of Monoclonal Antibody Against Human von Willebrand Factor Propeptide

To develop monoclonal antibodies that can specifically recognize human von Willebrand factor (VWF) propeptide (VWFpp) in plasma, and establish a rapid and reliable method for the detection of VWFpp antigen in plasma by using the double-antibody sandwich ELISA with the obtained anti-VWFpp monoclonal antibody. The recombinant human VWFpp (D1 and D2 regions) protein expressed in eukaryotic cells was used as immunogen to immunize BALB/c mice with routine method, so as to obtain clones of fusion cells. After screening and identification, hybridoma cell lines secreting monoclonal antibodies against VWFpp were selected, and then double-antibody sandwich ELISA assay was used to construct VWFpp antigen detection kit for the determination of VWFpp in human plasma. The levels of VWFpp antigen in plasma of 12 leukemia patients who underwent bone marrow transplantation were dynamically detected. Two hybridoma cell lines that can be subcultured continuously and secrete monoclonal antibodies against VWFpp were obtained and named SZ175 and SZ176 respectively. Identified by ELISA and Western blot, the antibodies could both specifically recognize VWFpp but couldn't recognize mature VWF (without propeptide). Based on the principle of double-antibody sandwich ELISA, monoclonal antibodies SZ175 and SZ176 were successfully made into a kit for detecting VWFpp antigen. The plasma VWFpp levels of leukemia patients before and after bone marrow transplantation were dynamically detected. The results showed that the plasma VWFpp levels of the patients after transplantation were significantly higher than those before transplantation. Two monoclonal antibodies against VWFpp were successfully prepared, and a double-antibody sandwich ELISA detection kit for VWFpp antigen was constructed, which provides a powerful tool for further study on the biological function of VWFpp, the clinical diagnosis and classification of von Willebrand disease (VWD), and the prognostic monitoring of endothelial injury-related diseases.

High-throughput thermal denaturation of tryptophanyl-tRNA synthetase combinatorial mutants reveals high-order energetic coupling determinants of conformational stability.

Landscape descriptions provide a framework for identifying functionally significant dynamic linkages in proteins but cannot supply details. Rate measurements of combinatorial mutations can implicate dynamic linkages in catalysis. A major difficulty is filtering dynamic linkages from the vastly more numerous static interactions that stabilize domain folding. The Geobacillus stearothermophilus (TrpRS) D1 switch is such a dynamic packing motif; it links domain movement to catalysis and specificity. We describe Thermofluor and far UV circular dichroism melting curves for all 16 D1 switch variants to determine their higher-order impact on unliganded TrpRS stability. A prominent transition at intermediate temperatures in TrpRS thermal denaturation is molten globule formation. Combinatorial analysis of thermal melting transcends the protein landscape in four significant respects: (i) bioinformatic methods identify dynamic linkages from coordinates of multiple conformational states. (ii) Relative mutant melting temperatures, δTM, are proportional to free energy changes. (iii) Structural analysis of thermal melting implicates unexpected coupling between the D1 switch packing and regions of high local frustration. Those segments develop molten globular characteristics at the point of greatest complementarity to the chemical transition state and are the first TrpRS structures to melt. (iv) Residue F37 stabilizes both native and molten globular states; its higher-order interactions modify the relative intrinsic impacts of mutations to other D1 switch residues from those estimated for single point mutants. The D1 switch is a central component of an escapement mechanism essential to free energy transduction. These conclusions begin to relate the escapement mechanism to differential TrpRS conformational stabilities.

Emerging Nosocomial Trichosporon asahii in a Tertiary Hospital in Southern Saudi Arabia

Background: Trichosporon species are pathogenic yeasts that colonize diverse human body sites and cause superficial in addition to invasive trichosporonosis, especially in immunosuppressed patients. The disease is an emerging health-care problem, as it has shown a noticeable increase in recent years. The leading risk factors are antibiotic use, invasive medical equipment, and chemotherapy. Informed evidence on the disease and its causal agent is needed. Objective: Presentation of clinical cases caused by Trichosporon sp. and identification of isolates recovered from various specimens in Aseer Central Hospital setting. Materials and Methods: The study analyzed fungal cultures (n = 20) from different clinical specimens submitted on a routine basis from hospital wards for general microbiological investigation at Aseer Central Hospital, Abha, Saudi Arabia (2021–2023). Mycologically proven trichosporonosis cases were initially identified by desktop methods, automated Vitek 2, and confirmed by sequence analysis of the ribosomal targets, namely the large-subunit RNA gene (D1–D2 region) and internal transcribed spacers 1 and 2 (ITS1 and ITS2 regions), patients were treated with fluconazole, and treatment outcomes were recorded. Results: Clinical features and culture results confirmed trichosporonosis caused by Trichosporon asahii with different clinical presentations. The majority of the cases were urinary tract (44%), followed by skin (28%) and respiratory tract (12%). All trichosporonosis cases responded to fluconazole. The neighbor-joining phylogenetic tree based on the rRNA regions and ITS and comparative analysis using GenBank reference data showed the association of our strains to the reference T. asahii clade, closely related to T. faecale and T. insectorum but different from other species. Conclusions: The study offered accurate species identification of the Trichosporon isolates by routine phenotypic tests as well as sequence-based techniques. Urinary tract infections were the most common manifestation of the infection, followed by skin and then respiratory tract infections. The cases of trichosporonosis responded well to fluconazole treatment. T. asahii is possibly underreported and should be considered in routine diagnostic investigations, notably in nosocomial infections.

Exploration of Multi-Gene DNA Barcode Markers to Reveal the Broad Genetic Diversity of Field Ticks (Acari: Ixodidae) in a Tropical Environment of Hainan Island, China

Ticks are hematophagous arthropods and obligate ectoparasites of humans and other animals. This study focused on the molecular discrimination of ticks in the tropical environment of Hainan according to multi-gene DNA barcode markers with the expectation of accurately distinguishing species. A total of 420 ticks, including 49 adult ticks, 203 nymphal ticks, and 168 larval ticks, were collected in the field, and the 49 adult ticks were identified as Rhipicephalus turanicus, Dermacentor marginatus, and Haemaphysalis longicornis. The mitochondrial 16S rRNA, ribosomal 28S rRNA D2, and ribosomal internal transcribed spacer 2 (ITS2) regions were used as DNA barcode markers to discriminate species. According to basic local alignment search tool analysis against the GenBank database, 16S rRNA positively identified ticks in the Rhipicephalus, Dermacentor, and Haemaphysalis genera; the 28S rRNA D2 region identified ticks in the Rhipicephalus and Dermacentor genera; and ITS2 identified ticks as D. marginatus. Pairwise sequence comparisons based on these three regions were visualized with a Sequence Demarcation Tool matrix. Substitution saturation tests using data analysis and molecular biology and evolution revealed little substitution saturation (Iss < Iss.c, p < 0.05) in the 16S rRNA region for the Haemaphysalis genus; 28S rRNA D2 region for the Rhipicephalus, Dermacentor, and Haemaphysalis genera; and ITS2 region for the Rhipicephalus and Dermacentor genera. Distinctive sequences for which it is difficult to obtain good matches with the sequences available in GenBank exist in the ticks of Hainan. Future studies should obtain complementary sequences to refine and update the database for the molecular characterization of ticks.

Diptacus rubuscolum Trinidad, Duarte and Navia (Eriophyoidea: Diptilomiopidae) on blackberry (Rubus ulmifolius Schott) in Mexico: occurrence, complementary description, and biology

Blackberry (Rubus ulmifolius Scott) is an economically important crop in Mexico. Eriophyoidea mites are among the most important arthropod pests affecting production of this crop worldwide. Despite this, reports on species diversity of Eriophyoidea on blackberry in Mexico are scarce. We sampled six orchards from two municipalities in the state of Michoacan, the most important region for blackberry production. Sampling was carried out monthly between May and November 2018. Only one species of eriophyoid mite was found, Diptacus rubuscolum Trinidad, Duarte and Navia. We conducted a detailed morphological study of this mite and a complementary description is presented. In addition, we obtained partial sequences of the mitochondrial cytochrome oxidase subunit I (mt COI), the 18S and 28S of the rRNA. The duration of each developmental stage was determined under controlled conditions (25 °C, 60% RH and 12:12 light regime). We confirmed the identity of D. rubuscolum by comparing partial sequences of the D2 region within the 28S rRNA from our samples and sequences retrieved from GenBank. Overall, morphological characteristics were in line with the type specimens. However, some differences on body shape and empodium structure were pointed out. The developmental stages of D. rubuscolum were egg, larva, nymphochrysalis, nymph, imagochrysalis and adult, with a mean duration of 75 (±10), 56 (±9), 21 (±7), 41 (±8), 19 (±6) and 114 (±12) hours respectively. The information produced provides insight and knowledge on the occurrence of this species on blackberry in Mexico as well as on original biological aspects.

Robotic-Assisted Transabdominal Inferior Retroperitoneal Approach: an Alternative to Cattell-Braash Maneuver

IntroductionThe Cattell-Braasch maneuver has been widely used to provide adequate exposure for aorto-caval space (ACS) since the 1960s. Given its requirement of complex visceral mobilization and significant physiological disturbance, we proposed a new alternative surgical technique to access ACS: the transabdominal inferior retroperitoneal approach (TIRA) with robotic assistance. MethodsPatients were placed in the Trendelenburg position, and the retroperitoneum was accessed from the iliac artery level and dissected toward the 3rd and 4th portion of the duodenum along the anterior surface of IVC and aorta. ResultsTIRA has been used in 5 consecutive patients at our institution whose tumors were located in the ACS below the level of SMA origin. The tumor sizes ranged from 1.7 to 5.6 cm. The median OR time was 192 min with a median EBL of 5 ml. Four out of the five patients passed flatus before or on post operative day (POD) 1, and the other patient passed flatus at POD2. The shortest length of stay was < 24 h, and the longest was 8 days due to pre-existing pain (the median length of stay was 4). ConclusionThe proposed robotic-assisted TIRA procedure is designed for tumors in the inferior portion of ACS, specifically for the tumor involving D3, D4, para-aortic, para-caval, and kidney regions. Since this approach does not involve organ mobilization and all the dissections are following an avascular plane, it can be easily adapted to laparoscopic and/or open surgery setting.

Crustal‐Scale Disharmonic Structural Pattern of West Junggar: Unveiling a Permian Indentation of Junggar Block Into Northern Kazakhstan Orocline

AbstractAs a crustal‐scale orocline in the Central Asian Orogenic Belt (CAOB), the Kazakhstan Orocline in particular its northern limb intervenes the Junggar Block and Chinese Altai. The West Junggar exhibits a structural pattern that is disharmonic from the major structure of the orocline. How and when such a structural complexity was formed remains poorly understood. To address this issue, we carried out detailed mapping, structural analysis and revision of geophysical data in three key areas of the West Junggar. It is shown that different arc chains experienced independent geological histories before they accreted with one another. Such an accretionary process generated the regional D1 deformation in late Carboniferous. Subsequent D2 shortening event produced variable superimposed structures, including crescent‐mushroom‐like folding in the Boshchekul‐Chingiz Arc and partitioned sinistral shearing of the West Karamay Unit. The synformal folding in the Zharma‐Saur Arc and crustal‐scale Erqis‐Zaysan sinistral strike‐slip shearing in late Permian—Triassic probably resulted from changing configuration of regional stress. Combined with previous data, we propose that the orogenic fabrics of arc chains and accretionary wedges were primarily related to shaping the basic structural framework of the Kazakhstan Orocline until late Carboniferous. The anticlockwise rotation of the northern limb of the orocline was associated with the indentation of the Junggar Block into the arc chains. Progressive indentation resulted in the first passive bending of arc chains followed by deformation partitioning into simple shear dominated transpression along the western margin of the Junggar Block and pure shear dominated transpression away from the indenter.

Revisiting the phylogeny of the family Miridae (Heteroptera: Cimicomorpha), with updated insights into its origin and life history evolution

Heteroptera is one of the most successfully adapted groups on Earth and can be observed in almost every environment. Within the evolution of heteropteran insects, Miridae show remarkable diversity (>11,700 spp.), accounting for a quarter of all Heteroptera. However, their phylogeny is still unclear, and no plausible theory for the driving force of their diversification has been established. In this work, we provide new suggestions for the phylogeny of Miridae using a larger dataset than previous studies. In addition, we suggest an alternative evolutionary history based on newly calibrated divergence dates for Miridae and its subordinate groups, and present probable factors of the family’s success in terms of species diversity. The entire dataset comprises 16 outgroups and 188 ingroup taxa including all seven known subfamilies and 37 out of 45 known tribes. Each species is aligned as 3,577 bp with six molecular loci (COI, 16S rRNA, 18S rRNA, 28S rRNA D3 region, H2A, and H3A). Among the molecular markers, we are the first to test histone genes (H2A, H3A) in Miridae. Our results raise the following points about phylogenetic relationships: i) The earliest group to diverge from Miridae was Monaloniini (Bryocorinae). ii) Bryocorinae and Cylapinae are polyphyletic, Deraeocorinae and Orthotylinae also rendered as non-monophyletic group. iii) Termatophylini and Coridromiini separated from Deraeocorinae and Orthotylinae respectively. iv) Four large tribes, Orthotylini, Phylini, Deraeocorini and Mirini are non-monophyletic. The results from our ancestral state reconstruction and divergence date estimation suggest the following: i) Miridae first diverged during the Late Jurassic (approx. 163.4 Mya), and the divergence dates of most subfamilies and tribes overlap with angiosperm radiation, which perhaps synergized their diversification. ii) Ancestral reconstruction results for Miridae reveal it to be predominantly phytophagous and diverge to oligophagy mainly in plant-tissue habitats, which could have allowed the mirids to select optimal tactics as plant-dwellers. iii) The common ancestor of Miridae originated among plant-dwellers mainly on Eudicots, and that tendency was largely maintained, but sporadic host shifts also occurred.