Tail Tip Research Articles

7940 Is Urocortin 2 A Mediator Of Stress-induced Suppression Of The Luteinizing Hormone Surge?

Abstract Disclosure: H.F. Bell: None. R.A. Carrasco: None. M.J. Kreisman: None. R.B. McCosh: None. K.M. Breen Church: None. Stress is thought to be a contributing factor to the reduction of reproductive function and infertility observed in many different species. Although this is a well-documented phenomenon, the neural mechanisms mediating the suppression of reproductive neuroendocrine activity in response to stress are not yet fully understood. The paraventricular nucleus (PVN) of the hypothalamus is an integral part of the neural response to stress. The PVN initiates activation of Corticotropin Releasing Hormone (CRH) neurons and, in addition to CRH, the PVN contains a population of neurons expressing Urocortin 2 (UCN2), a CRH-like family member, with high affinity for CRH receptor 2. In females, stress has been shown to inhibit both the pulse and surge modes of Luteinizing Hormone (LH) secretion. We have preliminary evidence that kisspeptin neurons, controlling either type of LH secretion, contain CRH receptor 2. In this study we tested the sufficiency of UCN2 to disrupt the estradiol-induced LH surge. Female mice were ovariectomized and given a high estradiol treatment to induce a circadian-timed LH surge. Two days later, UCN2 or Vehicle was administered into the lateral ventricle via ICV injection on the morning of the expected surge (10 AM). Blood and brains were collected in the evening (6 PM), at the time of the expected surge. Whole blood from the tail tip was measured by LH ELISA and a surge was defined as > 0.5 ng/mL. A majority of saline-treated animals expressed an LH surge (8 of 9 females). In contrast, 0% of UCN2-treated animals surged (n=7), suggesting that UCN2 impaired the positive feedback response to estradiol. Additionally, immunohistochemistry was performed to observe colocalization of c-Fos within Kisspeptin neurons in the rostral periventricular region of the third ventricle (RP3V) which are responsible for surge generation. Lower Kisspeptin/C-Fos co-expression in animals treated with UCN2 demonstrated the efficacy of UCN2 to impair estradiol-induced activation of RP3V Kisspeptin neurons which are critical for LH surge generation. These results support the hypothesis that UCN2 signaling plays a mediatory role in the stress-induced reduction of ovulatory function in the female mouse. Presentation: 6/1/2024

Phosphatidylserine-blocking nanoparticles inhibit thrombosis without increased bleeding in mice

BackgroundPhosphatidylserine (PS) is a procoagulant phospholipid enriched on surfaces of activated vascular cells including platelets, endothelium, monocytes, and microvesicles. As a molecular driver of thrombosis accessible to drug blockade, PS is an attractive pharmacologic target for modulating thrombogenesis, with potentially reduced bleeding risk compared to anticoagulant and antiplatelet therapies. ObjectivesTest antithrombotic capabilities of a liposomal formulation, Zn-dipicolylamine cyanine-3[22,22]/1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (molar ratio, 3:97), designated as DPAL, which we previously described binds selectively to PS-enriched cell surfaces, compared with effects on bleeding, in mouse models. MethodsPS-dependent DPAL binding to human and murine platelets was tested in vitro. Thrombosis and bleeding after DPAL intravenous administration were tested in C57Bl/6J mice following FeCl3 carotid arterial injury and tail tip amputation, respectively. Incorporation in hemostatic clots was investigated in the cremaster muscle laser injury model. Toxicity was tested by direct exposure to human endothelial cell cultures. ResultsDPAL bound agonist-stimulated, PS-positive human and murine platelets, blocked by Annexin V or Ano6 deletion, which ablate PS exposure. DPAL prolonged prothrombin time, but did not prevent thrombin-induced fibrinogen receptor activation or aggregation, nor alter blood cell counts including platelets. Following arteriolar laser injury, DPAL bound wound surfaces and edges without destabilizing plugs. DPAL dose-dependently blocked FeCl3-induced arterial thrombosis but did not substantially increase bleeding, or induce endothelial cell death. ConclusionDPAL reduces thrombogenesis with minimal effects on bleeding in mouse models via selective binding to PS. DPAL may support novel approaches to modulate pathogenic thrombin generation with improved safety profiles in multiple contexts.

A new four-pored Amphisbaena Linnaeus, 1758 (Amphisbaenia, Amphisbaenidae) from the north of Espinhaço Mountain Range, Brazil.

A new species of Amphisbaena is described from the north of Espinhaço Mountain Range, municipality of Caetité, state of Bahia, Brazil. Amphisbaenaamethysta sp. nov. can be distinguished from its congeners by the following combination of characters: (1) snout convex in profile, slightly compressed not keeled; (2) pectoral scales arranged in regular annuli; (3) four precloacal pores; (4) distinct cephalic shields; (5) 185-199 dorsal half-annuli; (6) 13-16 caudal annuli; (7) conspicuous autotomic site between 4th-6th caudal annuli; (8) 16-21 dorsal and ventral segments at midbody; (9) 3/3 supralabials; (10) 3/3 infralabials; and (11) smooth and rounded tail tip. The new species is the 71st species of genus with four precloacal pores, and the 22nd species from the Caatinga morphoclimatic domain. The identification of Amphisbaenaamethysta sp. nov. indicates that the reptile fossorial fauna in the Espinhaço Mountain Range region is far from being completely known and that it may harbour a much greater diversity of endemic taxa.

Tissue-specific RNA-seq defines genes governing male tail tip morphogenesis in C. elegans.

Caenorhabditis elegans males undergo sex-specific tail tip morphogenesis (TTM) under the control of the DM-domain transcription factor DMD-3. To find genes regulated by DMD-3, we performed RNA-seq of laser-dissected tail tips. We identified 564 genes differentially expressed (DE) in wild-type males versus dmd-3(-) males and hermaphrodites. The transcription profile of dmd-3(-) tail tips is similar to that in hermaphrodites. For validation, we analyzed transcriptional reporters for 49 genes and found male-specific or male-biased expression for 26 genes. Only 11 DE genes overlapped with genes found in a previous RNAi screen for defective TTM. GO enrichment analysis of DE genes finds upregulation of genes within the unfolded protein response pathway and downregulation of genes involved in cuticle maintenance. Of the DE genes, 40 are transcription factors, indicating that the gene network downstream of DMD-3 is complex and potentially modular. We propose modules of genes that act together in TTM and are co-regulated by DMD-3, among them the chondroitin synthesis pathway and the hypertonic stress response.

On the autotomy in Pattonomys semivillosus (Rodentia: Echimyidae) and Calomys hummelincki (Rodentia: Cricetidae) in Colombia

Tail autotomy is rare among mammals, mainly observed in rodents. This note reports two new cases in the Colombian Speckled Tree Rat (Pattonomys semivillosus) and Hummelinck’s vesper mouse (Calomys hummelincki). P. semivillosus displays real autotomy, dropping its entire tail when grabbed, akin to lizard behavior. C. hummelincki exhibits fake autotomy, shedding only the tail tip, exposing the last eight vertebrae. These observations add to the limited knowledge of autotomy in rodents, increasing the known species count to 49.

Structures of Mature and Urea-Treated Empty Bacteriophage T5: Insights into Siphophage Infection and DNA Ejection.

T5 is a siphophage that has been extensively studied by structural and biochemical methods. However, the complete in situ structures of T5 before and after DNA ejection remain unknown. In this study, we used cryo-electron microscopy (cryo-EM) to determine the structures of mature T5 (a laboratory-adapted, fiberless T5 mutant) and urea-treated empty T5 (lacking the tip complex) at near-atomic resolutions. Atomic models of the head, connector complex, tail tube, and tail tip were built for mature T5, and atomic models of the connector complex, comprising the portal protein pb7, adaptor protein p144, and tail terminator protein p142, were built for urea-treated empty T5. Our findings revealed that the aforementioned proteins did not undergo global conformational changes before and after DNA ejection, indicating that these structural features were conserved among most myophages and siphophages. The present study elucidates the underlying mechanisms of siphophage infection and DNA ejection.

A Look Inside-Histopathological Examinations of Different Tail Tip Lesions in Dairy Cows.

Feedlot cattle are frequently affected by inflammation and necrosis of the tail tips, resulting in impeded animal welfare and economic losses. In a recent study, it was demonstrated that dairy cows are also affected by different lesions of the tail tip, including alopecia, annular constrictions, crusting, scaling and swelling. Despite the frequent occurrence of these lesions, the underlying etiology and pathomechanisms are unclear to date. To gain insight into this malady, we histopathologically examined 16 tail tips of slaughtered dairy cows, representing the entire range of different lesions. While macroscopically alopecic areas were characterized by the formation of granulation tissue in the dermis with an inconspicuous epidermis, we found not only dermal granulation tissue but also purulent-necrotizing inflammation with the breakdown of the basement membrane in encrusted lesions. Interestingly, in some cases, we found areas of coagulation necrosis of the epidermal and dermal layers in the crusts. Tails with macroscopical scaling were affected by ortho- and/or parakeratotic hyperkeratosis, and, in tails with macroscopical swelling, we observed a low-protein edema of the central longitudinal connective tissue of the tail. We conclude that the observed lesions might be caused by ischemia of the skin in the distal parts of the tail.

P-154 Structural changes of the human sperm tail during the early post-fertilization

Abstract Study question What happens to the sperm tails that are no longer needed after the goal of fertilization has been achieved? Summary answer The sperm tail lost its fibrous sheath and changed to a coiled morphology with a whisker root-like structure at the tip during the early post-fertilization. What is known already The sperm tail has been thought to play only a role in allowing the head and neck to reach the oocyte. The flagellum is found in the mammalian sperm tail and consists of a middle, main, and terminal part. It also contains an axon composed of two central microtubules surrounded by nine peripheral microtubule doublets (9d + 2s microtubule pattern) surrounded by an accessory structure called the fibrous sheath (FS), but few studies have been conducted on the post-fertilization fate of the mammalian sperm tail. Study design, size, duration To investigate sperm tail structure from the pronuclear stage to the second cleavage, we used abnormally fertilized oocytes derived from either conventional in vitro fertilization (cIVF, n = 41) or intracytoplasmic sperm injection (ICSI, n = 11). In this study, abnormally fertilized oocytes were collected between March 2022 and November 2023 and subjected to immunofluorescence analysis and field emission scanning electron microscopy (FE-SEM) analysis. Participants/materials, setting, methods Under ethical approval, abnormally fertilized oocytes were donated by infertile couples undergoing cIVF or ICSI cycles at the Yamashita Shonan Yume Clinic in Japan. Informed consent was obtained from all participants. Immunofluorescence assay was performed to analyze sperm tail morphology using antibodies against H3K27me3, α-tubulin, and AKAP4. FE-SEM analysis was performed to detect the 9d + 2s structure characteristic of the sperm tail axon. Main results and the role of chance Tri-pronuclear zygotes derived from conventional in vitro fertilization (cIVF-derived 3PN) had two male pronuclei, whereas those derived from intracytoplasmic sperm injection (ICSI-derived 3PN) had one. Each zygote had the same number of coiled microtubule structures as the male pronucleus, and the 9d + 2s structure characteristic of the axon was identified, suggesting a sperm tail. No difference in sperm tail shape was observed between ICSI-derived 3PN and cIVF-derived 3PN. AKAP4, which is specifically localized in the FS associated with the principal piece of the flagellum, was observed in sperm tails attached to the zona pellucida but not in those in the oocyte, suggesting that the FS of the flagellum is lost before the pronuclear stage, leaving the microtubules of the axonemes exposed. The sperm tail was attached to one of the spindle poles during the first cleavage of abnormally fertilized oocytes, but no differences in the structure of the spindle pole with or without the sperm tail were observed. The tip of the sperm tail was branched like a “whisker root”, suggesting that the sperm tail was degraded from the end, but the degree of tip branching did not differ from the pronuclear stage to the second cleavage. Limitations, reasons for caution Normal fertilized oocytes were not used in this study because they are used for clinical treatment. The cell biological significance of sperm tail branching is currently unknown and requires further experimentation. Wider implications of the findings These findings shed light on the processes that occur during human fertilization, suggest new roles for the sperm tail, and may have implications for assisted fertilization approaches in the future. Trial registration number not applicable

A pilot study: tail tip lesions in dairy cows – an unnoticed animal welfare issue?

Abstract. The welfare of dairy cows is becoming increasingly important. While diseases like mastitis and lameness are common ailments, injuries to the tail tip go largely unnoticed. This study aimed to investigate whether tail tip lesions, which are mostly described in beef cattle, also occurred on n=5 dairy farms, along with determining what type and at what frequency. The study consisted of two phases. During the first part of the study, tail tips of 78 dairy cows were examined over a period of 6 months; based on these results, we developed a training card on tail tip lesions in dairy cows, which was used in part two of the study to train further examiners to inspect four more flocks. In total, we collected n=3587 tail records from n=513 Holstein and n= 128 Simmental dairy cows. The overall frequency regarding all types of lesions ranged between 84.0 % (±2.0) and 94.1 % (±1.8) in Holstein herds and between 97.0 % (±2.2) and 99.0 % (±2.2) in Simmental herds. To our knowledge, this is the first investigation of tail tip lesions in German dairy cows. We concluded that tail tip lesions might be a frequent yet unnoticed condition in German dairy cows.

Morphologically defined substages of tail morphogenesis in C. elegans males.

Sex-specific morphogenesis occurs in Caenorhabditis elegans in the vulva of the hermaphrodite and in the male tail during the last larval stage. Temporal progression of vulva morphogenesis has been described in fine detail. However, a similar precise description of male tail morphogenesis was lacking. We here describe morphogenesis of the male tail at time points matching vulva development with special focus on morphogenesis of the tail tip. Using fluorescent reporters, we follow changes in cell shapes, cell fusions, nuclear migration, modifications in the basement membrane, and formation of a new apical extracellular matrix at the end of the tail. Our analysis answers two open questions about tail tip morphogenesis (TTM) by showing that one of the four tail tip cells, hyp11, remains largely separate, while the other cells fully fuse with each other and with two additional tail cells to form a ventral tail syncytium. This merger of cells begins at the apical surface early during TTM but is only completed toward the end of the process. This work provides a framework for future investigations of cell biological factors that drive male tail morphogenesis.

An arabinogalactan isolated from Cynanchum atratum promotes lymphangiogenesis and lymphatic vessel remodeling to alleviate secondary lymphedema

Secondary lymphedema is a chronic and incurable disease lacking satisfactory therapeutic drugs. It primarily results from lymphatic vessel dysfunction resulting from factors such as tumor-related surgery, injury, or infection. Promoting lymphangiogenesis and lymphatic vessel remodeling is crucial for restoring tissue fluid drainage and treating secondary lymphedema. In this study, we discovered that the oral administration of a type-II arabinogalactan (CAPW-1, molecular weight: 64 kDa) significantly promoted lymphangiogenesis and alleviated edema in mice with secondary lymphedema. Notably, the tail diameter of the CAPW-1200 group considerably decreased in comparison to that of the lymphedema group, with an average diameter difference reaching 0.98 mm on day 14. CAPW-1 treatment also reduced the average thickness of the subcutaneous area in the CAPW-1200 group to 0.37 mm (compared with 0.73 mm in the lymphedema group). It also facilitated the return of injected indocyanine green (ICG) from the tail tip to the sciatic lymph nodes, indicating that CAPW-1 promoted lymphatic vessel remodeling at the injury site. In addition, CAPW-1 enhanced the proliferation and migration of lymphatic endothelial cells. This phenomenon was associated with the activation of the toll-like receptor 4 (TLR4)/nuclear factor-κB (NF-κB) signaling pathway, thereby promoting the expression of vascular endothelial growth factor-C (VEGF-C), which can be abolished using a TLR4 antagonist. Despite these findings, CAPW-1 did not alleviate the symptoms of lymphedema or restore lymphatic drainage in VEGFR3flox/flox/Prox1-CreERT2 mice. In summary, CAPW-1 alleviates secondary lymphedema by promoting lymphangiogenesis and lymphatic vessel remodeling through the activation of the TLR4/NF-κB/VEGF-C signaling pathway, indicating its potential as a therapeutic lymphangiogenesis agent for patients with secondary lymphedema.

Comparison of Thermal and Mechanical Pain Testing Modalities in Sprague Dawley and Fischer 344 Rats (Rattus norvegicus).

While rodents are used extensively for studying pain, there is a lack of reported direct comparisons of thermal and mechanical pain testing methods in rats of different genetic backgrounds. Understanding the range of interindividual variability of withdrawal thresholds and thermal latencies based on these testing methods and/or genetic background is important for appropriate experimental design. Testing was performed in two common rat genetic backgrounds: outbred Sprague-Dawley (SD) and inbred Fischer 344 (F344). Male and female, 10- to 14-wk-old F344 and SD rats were used to assess withdrawal thresholds in 3 different modalities: the Randall-Selitto test (RST), Hargreaves test (HT), and tail flick test (TFT). The RST was performed by using an operator-controlled handheld instrument to generate a noxious pressure stimulus to the left hind paw. The HT and the TFT used an electronically controlled light source to deliver a noxious thermal stimulus to the left hind paw or tail tip, respectively. Rats of each sex and genetic background underwent one type of test on day 0 and day 7. Withdrawal thresholds and thermal latencies were compared among tests. No significant differences were observed. Our findings can serve as a guide for researchers considering these nociceptive tests for their experiments.

Morphological and molecular notes on root lesion nematode Pratylenchus coffeae and Pratylenchus neglectus (Nematoda: Pratylenchidae) from Pakistan

During a survey, a root lesion nematode Pratylenchus coffeae was recovered from the soil associated with Plantago ovata plantation in Sindh, Pakistan. The first report of P. coffeae from soil and root of Plantago ovata was examined morphologically and molecularly. The Pakistani population had few geographical variation and is characterized by body length (L = 360–450; a = 18.0–22.5, c = 7.2–16.1, c' = 1.7–3.1, V% = 72–80), a labial region with 2 annuli, a lateral field with 6 incisures, stylet 16–17 μm long with rounded knobs, spermatheca reduced or absent, post vulval uterine sac 28–35 μm long, and tail tip smooth, rounded, and containing 19–23 annuli ventrally. Morphometric and molecular features of 18 S rDNA, and mitochondrial cytochrome oxidase subunit 1 (cox1) genes are characterized. Phylogenetic analysis using cox1 of mtDNA place it close to P. coffeae, and P. speijeri with 1.00 posterior probability values. Measurements, line illustrations, and LM pictures for P. coffeae are given. Further, the detailed morphological, molecular, Discriminant analysis (DA) revealed differences between the P. coffeae and the close species morphologically and molecularly. Besides, P. neglectus verified using morphology and molecular characters. The nblast result of 18 S rDNA and cox1 of mtDNA showed 100 % similarity with the same species. Phylogenetic analysis using 18 S rDNA and cox1 placed the Pakistanis population of P. neglectus together with the other P. neglectus in a clade with 1.00 posterior probability. Population study of P. neglectus associated with grapevines (Vitis vinifera L.) revealed that this species had the most prominence value in Abbottabad of Khyber Pakhtun Khwa Province of Pakistan.

Structural mechanism of bacteriophage lambda tail’s interaction with the bacterial receptor

Bacteriophage infection, a pivotal process in microbiology, initiates with the phage’s tail recognizing and binding to the bacterial cell surface, which then mediates the injection of viral DNA. Although comprehensive studies on the interaction between bacteriophage lambda and its outer membrane receptor, LamB, have provided rich information about the system’s biochemical properties, the precise molecular mechanism remains undetermined. This study revealed the high-resolution cryo-electron microscopy (cryo-EM) structures of the bacteriophage lambda tail complexed with its irreversible Shigella sonnei 3070 LamB receptor and the closed central tail fiber. These structures reveal the complex processes that trigger infection and demonstrate a substantial conformational change in the phage lambda tail tip upon LamB binding. Providing detailed structures of bacteriophage lambda infection initiation, this study contributes to the expanding knowledge of lambda-bacterial interaction, which holds significance in the fields of microbiology and therapeutic development.

Variations in salivary oxytocin and eye caruncle temperature indicate response to environmental enrichment material in fattening pigs

Conventional pigs are typically raised under intensive conditions, which often do not meet pigs’ basic needs regarding behavioural expression and cognitive challenge, likely threatening their welfare. Thus, enabling the experience of positive affective states and, consequently, positive welfare should be encouraged. We investigated the potential of salivary oxytocin (sOXT) and eye (i.e., lachrymal caruncle region) temperature measured by infrared thermography as indicators of environmental enrichment material valuing in 32 conventional Yorkshire × Landrace pigs (110-day-old, mean ± SD body weight: 80.9±7.6 kg/pig) housed in 8 identical pens (3 female and 1 castrated male/pen; 3.3 m2/pig). Pigs were subjected to two contexts/days: fulfilment of expectation for fresh straw (i.e., delivery of 200 g of fresh straw/pig) and violation of expectation (i.e., no straw delivered). Additionally, based on footage of individual pigs from −30 min to +30 min relative to the moment when an experimenter left the pen after straw had been delivered or not (reference moment), we labelled behaviours more extensively investigated as measures of affective states (body posture; locomotor-rotational play, LOC; social play, SOC; aggression; tail motion; tail posture; tail tip lateralisation). Individual saliva samples were collected at −10 min and +20 min for analysis of sOXT and cortisol (sCORT), and individual thermograms were recorded at −10 min, −5 min and +10 min relative to the reference moment. Pigs tended to move their tail more often and tended to display more aggressions following straw provision. Furthermore, pigs displayed more playful events after the reference moment than before this event, irrespective of straw availability. However, pigs were mostly inactive (i.e., sitting or lying down) across contexts and no clear behavioural differences between contexts were detected. Similarly, we found no clear differences in sCORT concentrations between contexts. Alternatively, eye caruncle temperature and sOXT concentrations were higher on days when straw was provided. Our results suggest that salivary oxytocin and lachrymal caruncle temperature can be useful non-invasive tools to detect short-term responses to non-social stimuli in fattening pigs. Further research is needed and this study supports continuous development and validation of measures of affective state in domesticated animals.

First report of the damaging pest Meloidogyne enterolobii parasitizing mulberry (Morus alba L.) in Vietnam.

Mulberry (Morus alba L.) is highly important crop in Vietnam, playing a key role in the country's economy through sericulture, food supply, pharmaceuticals, and beverage industries (Nguyen et al., 2018; Rohela et al., 2020). Recently, many mulberry-growing areas in Lam Dong, Vietnam have reported severe symptoms associated with nematode infection, including yellowing leaves, stunted growth, and severe root galling, leading to a significant decline in mulberry productivity. From April to December 2022, twenty soil and root samples from mulberry-growing areas in Lam Dong (Da Teh: 11°28'48.11"N; 107°28'23.74"E elevation: 133m; Lam Ha 11°48'25.13"N; 108°14'7.13"E elevation: 848m) were collected to uncover the presence of Meloidogyne enterolobii parasitizing mulberry in Vietnam. One nematode population was randomly selected for characterizing in this study among analyzed nematode populations. Females were extracted from heavily galled roots (Fig. S1) from a single mulberry tree in Lam Dong, Vietnam, using a needle and forceps (Subbotin et al., 2021). The perineal patterns of adult females (n = 10) have an oval shape, with clearly visible phasmids, along with a prominently high and squared dorsal arch. The striae are smooth and coarse, while the perivulval region remains devoid of striae. The lateral lines appear indistinct, and the tail tip is easily observable. Morphometric measurements were as follows: body length = 585 ± 78 (464-724) µm, body width = 367 ± 75 (271-529) µm, neck length = 221.5 ± 30.7 (167-269.6) µm, stylet length = 13.1 ± 1.2 (11.4-15.1) µm, vulva-slit length 16.3±2.3 (10.4-18) µm, vulva-anus distance = 16.8±3.0 (11.4-18) µm, anus-tail tip distance = 10.3±2.1 (6.9-14.2) µm, interphasmidial distance = 15.9 ± 3.7 (10.3-23.4) µm. The morphology of this nematode population is highly in agreement with the original description of M. enterolobii (Yang & Eisenback, 1983). This population was also identified using the D2-D3 of 28S rRNA and 18S rRNA (Powers et al., 2017; Subbotin et al., 2006) regions. The D2-D3 of 28S rRNA sequences from this study (accession numbers: OR889633) exhibited 99.5-99.8% similarity to the sequences of M. enterolobii from GenBank (accession numbers: OR214950 and ON496981). While the 18S rRNA sequences (accession numbers: OR896547) showed 99.2-99.3% similarity to the sequences of M. enterolobii from GenBank (accession numbers: MZ955995, MZ531901, and MW488150). To carry out Koch's postulates, 2000 J2s from collected M. enterolobii egg masses (initial population) were inoculated on two-month-old plantlets of mulberry (n = 6), planted on 2L pots within a screenhouse, non-inoculated plantlets (n=6) served as negative controls. After 90 days post-inoculation, nematode reproduction factors (RF = final density (nematodes were extracted from the whole root system and corresponding soil samples (Subbotin et al., 2021)) / initial population) and root damage symptoms were evaluated. The inoculated plantlets exhibited consistent yellowing leaves, stunting, and root galling symptoms (Fig. S1), mirroring observations from the field, with an average RF of 11.5. Control plants displayed no symptoms. Root-knot nematodes extracted from the roots were identified as M. enterolobii through molecular analyses of D2-D3 of 28S and 18S rRNA regions (GenBank accession numbers: OR889634 (D2-D3 of 28S) and OR896548 (18S)), thereby confirming that mulberry acts as a host for M. enterolobii. Currently, this nematode has been reported to be associated with two different host plants, including guava (Trinh et al., 2022) and pomelo (Le et al., 2023). Our discovery marks the first documented case of Meloidogyne enterolobii parasitizing mulberry in Vietnam. While the impact on mulberry productivity remains to be really important for sericulture food supply, pharmaceuticals, and beverage industries; the aggressive nature of M. enterolobii, as observed in the field and confirmed by the screenhouse tests, raises concerns about potential economic losses in mulberry production. Therefore, further investigations are needed to assess the extent of M. enterolobii infestation in mulberry orchards and to develop effective control measures to safeguard the sustainability of mulberry cultivation in Vietnam.

Description of Pauronemoides siddiqii n. gen., n. sp. (Tylenchomorpha, Paurodontidae), a morphological and molecular phylogenetic study

Summary Three populations of Pauronemoides siddiqii n. gen., n. sp. were recovered from three different locations in northwestern and northern Iran from bark and soil samples. The new genus was described and illustrated based upon morphological and molecular characteristics. Pauronemoides n. gen. belongs to the family Paurodontidae and is characterised by the medium-sized body, asymmetrical knobs in stylet base, stem-like extension into intestine, post-vulval uterine sac (PUS) absent or rudimentary, different tail tip shape between sexes, and bursa enveloping tail or reaching to the tail end. The type species, Pauronemoides siddiqii n. gen., n. sp., is mainly characterised by 752-1281 μm body length in females and 660-1085 μm in males, six equally-sized lips, stylet 9-11 μm long with asymmetrical knobs, six incisures in lateral fields forming five bands, three of which (two outers and one central) are wider, and variably lengthened stem-like extension projecting into the intestine. Vulva situated posteriorly (V = 87.7-91.7%), PUS absent. Conical tail with widely rounded end, having a small mucron at tip in females, males with conical tail ending to a narrower tip and bursa enveloping tail completely. The type species of the new genus was compared with all known species of the genera Prothallonema, Paurodontus, Paurodontoides and Paurodontella. Pauronemoides siddiqii n. gen., n. sp. resembles Prothallonema macrocellum and P. mucronatum by having the bulb extension penetrating into the intestine, dimorphism in tail shape of both sexes and bursa enveloping tail, and Paurodontella iranica by tail shape dimorphism between sexes and bursa ending near tail tip. Therefore, the above-mentioned species are transferred to the new genus as Pauronemoides macrocellus n. gen., n. comb. (= Prothallonema macrocellum), P. mucronatus n. gen., n. comb. (= Prothallonema mucronatum) and P. iranicus n. gen., n. comb. (= Paurodontella iranica). In addition to morphological comparisons, molecular phylogenetic analyses were performed using two sequences of small subunit (SSU), and three partial sequences of D2-D3 expansion segments of the large subunit (LSU) rDNA, and the resulting topologies were discussed.

EVALUATION OF THE HAEMOSTATIC EFFECT OF PANCHAVALKALA KASHAYA AND AQUEOUS EXTRACT IN WISTAR RATS

The effect of Panchavalkala group of drugs on blood coagulation has not been scientifically tested despite it has mentioned in Ayurveda by Acharya Sushruta. The current study investigates the effect of Panchavalkala kashaya and Panchavalkala aqueous extracts on Wistar rats by tail tip amputation method. Amount of bleeding and Haemostatic time is observed immediately after tail tip amputation in heparinised and non- heparinised groups. The data was analysed using Kruskal Wallis test. In non-heparinized category of Wistar rats proved that both Panchavalkala kashaya and Panchavalkala aqueous extract have highly significant haemostatic effect in the test animals. Panchavalkala aqueous extract was more effective in reducing haemostatic time as well as the amount of bleeding in heparinised Wistar rats when compared to Panchavalkala kashaya, which did not show any significant haemostatic effect in the heparinized test animals.

Phenotypic analysis of ataxia in spinocerebellar ataxia type 6 mice using DeepLabCut

This study emphasizes the benefits of open-source software such as DeepLabCut (DLC) and R to automate, customize and enhance data analysis of motor behavior. We recorded 2 different spinocerebellar ataxia type 6 mouse models while performing the classic beamwalk test, tracked multiple body parts using the markerless pose-estimation software DLC and analyzed the tracked data using self-written scripts in the programming language R. The beamwalk analysis script (BAS) counts and classifies minor and major hindpaw slips with an 83% accuracy compared to manual scoring. Nose, belly and tail positions relative to the beam, as well as the angle at the tail base relative to the nose and tail tip were determined to characterize motor deficits in greater detail. Our results found distinct ataxic abnormalities such as an increase in major left hindpaw slips and a lower belly and tail position in both SCA6 ataxic mouse models compared to control mice at 18 months of age. Furthermore, a more detailed analysis of various body parts relative to the beam revealed an overall lower body position in the SCA684Q compared to the CT-longQ27PC mouse line at 18 months of age, indicating a more severe ataxic deficit in the SCA684Q group.

Cryo-EM structure of flagellotropic bacteriophage Chi

The flagellotropic bacteriophage χ (Chi) infects bacteria via the flagellar filament. Despite years of study, its structural architecture remains partly characterized. Through cryo-EM, we unveil χ's nearly complete structure, encompassing capsid, neck, tail, and tail tip. While the capsid and tail resemble phage YSD1, the neck and tail tip reveal new proteins and their arrangement. The neck shows a unique conformation of the tail tube protein, forming a socket-like structure for attachment to the neck. The tail tip comprises four proteins, including distal tail protein (DTP), two baseplate hub proteins (BH1P and BH2P), and tail tip assembly protein (TAP) exhibiting minimal organization compared to other siphophages. Deviating from the consensus in other siphophages, DTP in χ forms a trimeric assembly, reducing tail symmetry from 6-fold to 3-fold at the tip. These findings illuminate the previously unexplored structural organization of χ's neck and tail tip.