Crystal structure of Middle East respiratory syndrome coronavirus helicase.

Middle East respiratory syndrome coronavirus (MERS-CoV) remains a threat to public health worldwide; however, effective vaccine or drug against CoVs remains unavailable. CoV helicase is one of the three evolutionary most conserved proteins in nidoviruses, thus making it an important target for drug development. We report here the first structure of full-length coronavirus helicase, MERS-CoV nsp13. MERS-CoV helicase has multiple domains, including an N-terminal Cys/His rich domain (CH) with three zinc atoms, a beta-barrel domain and a C-terminal SF1 helicase core with two RecA-like subdomains. Our structural analyses show that while the domain organization of nsp13 is conserved throughout nidoviruses, the individual domains of nsp13 are closely related to the equivalent eukaryotic domains of Upf1 helicases. The most distinctive feature differentiating CoV helicases from eukaryotic Upf1 helicases is the interaction between CH domain and helicase core.

Newborn Screening for Hemoglobinopathies and Red Cell Enzymopathies in Tripura State: A Malaria-Endemic State in Northeast India.

Hemoglobinopathies are a group of inherited single gene disorders. There are reports on hemoglobin (Hb) variants identified in the tribal and non-tribal populations of Tripura State in northeastern India. This study aimed to determine the spectrum of hemoglobinopathies and enzymopathies by newborn screening in Tripura State and assess the extent of neonatal jaundice. A total of 2400 cord blood samples were collected and analyzed by high performance liquid chromatography (HPLC). Further confirmation of any abnormal HPLC was done by DNA analysis. The samples were also screened for deficiency of enzymopathies, glucose-6-phosphate dehydrogenase (G6PD) deficiency and pyruvate kinase. Of 2400 cord blood samples screened, 225 (9.3%) were Hb E (HBB: c.79G>A) heterozygotes, 80 (3.3%) were Hb E homozygotes and one carried Hb E-ß-thalassemia (ß-thal). Other Hb abnormalities were also detected including 15 Hb S (HBB: c.20A>T) heterozygotes, two Hb D-Punjab (HBB: c.364G>C) heterozygotes and two compound heterozygotes for Hb D-Punjab and Hb E. Of the 80 homozygous Hb E babies, four were non-tribal and 76 babies were tribal, and 225 patients carried Hb E trait, 141 were tribal, while 84 were non-tribal. Of 40 G6PD deficient babies identified, 13 had coinherited Hb E and two babies had pyruvate kinase deficiency. α Genotyping was performed in 162 affected babies, 50 of them carried α gene deletions. Newborn screening programs for Hb E, other hemoglobinopathies and G6PD deficiency must be encouraged in the malaria-endemic northeastern region of India. Drug-induced hemolysis can also be avoided by screening for G6PD deficiency at birth.

Stress-stiffening-mediated stem-cell commitment switch in soft responsive hydrogels.

Bulk matrix stiffness has emerged as a key mechanical cue in stem cell differentiation. Here, we show that the commitment and differentiation of human mesenchymal stem cells encapsulated in physiologically soft (∼0.2-0.4 kPa), fully synthetic polyisocyanopeptide-based three-dimensional (3D) matrices that mimic the stiffness of adult stem cell niches and show biopolymer-like stress stiffening, can be readily switched from adipogenesis to osteogenesis by changing only the onset of stress stiffening. This mechanical behaviour can be tuned by simply altering the material's polymer length whilst maintaining stiffness and ligand density. Our findings introduce stress stiffening as an important parameter that governs stem cell fate in a 3D microenvironment, and reveal a correlation between the onset of stiffening and the expression of the microtubule-associated protein DCAMKL1, thus implicating DCAMKL1 in a stress-stiffening-mediated, mechanotransduction pathway that involves microtubule dynamics in stem cell osteogenesis.

Permanent uncoupling of male-specific CYP2C11 transcription/translation by perinatal glutamate.

Perinatal exposure of rats and mice to the typically reported 4mg/g bd wt dose of monosodium glutamate (MSG) results in a complete block in GH secretion as well as obesity, growth retardation and a profound suppression of several cytochrome P450s, including CYP2C11, the predominant male-specific isoform--all irreversible effects. In contrast, we have found that a lower dose of the food additive, 2mg/g bd wt on alternate days for the first 9days of life results in a transient neonatal depletion of plasma GH, a subsequent permanent overexpression of CYP2C11 as well as subnormal (mini) GH pulse amplitudes in an otherwise normal adult masculine episodic GH profile. The overexpressed CYP2C11 was characterized by a 250% increase in mRNA, but only a 40 to 50% increase in CYP2C11 protein and its catalytic activity. Using freshly isolated hepatocytes as well as primary cultures exposed to the masculine-like episodic GH profile, we observed normal induction, activation, nuclear translocation and binding to the CYP2C11 promoter of the GH-dependent signal transducers required for CYP2C11 transcription. The disproportionately lower expression levels of CYP2C11 protein were associated with dramatically high expression levels of an aberrant, presumably nontranslated CYP2C11 mRNA, a 200% increase in CYP2C11 ubiquitination and a 70-80% decline in miRNAs associated, at normal levels, with a suppression of CYP2C expression. Whereas the GH-responsiveness of CYP2C7 and CYP2C6 as well as albumin was normal in the MSG-derived hepatocytes, the abnormal expression of CYP2C11 was permanent and irreversible.

Irreversible perinatal imprinting of adult expression of the principal sex-dependent drug-metabolizing enzyme CYP2C11.

We proposed to determine whether, like other sexual dimorphisms, drug metabolism is permanently imprinted by perinatal hormones, resulting in its irreversible sex-dependent expression. We treated newborn male rats with monosodium glutamate (MSG), a total growth hormone (GH) blocker, and, using cultured hepatocytes, examined expression of adult CYP2C11, the predominant cytochrome-P450 expressed only in males, as well as the signal transduction pathway by which episodic GH solely regulates the isoform's expression. In addition, adolescent hypophysectomized (hypox) male rats served as controls in which GH was eliminated after the critical imprinting period. Whereas renaturalization of the masculine episodic GH profile restored normal male-like levels of CYP2C11, as well as CYP2C12, in hepatocytes from hypox rats, the cells derived from the MSG-treated rats were completely unresponsive. Moreover, GH exposure of hepatocytes from hypox rats resulted in normal induction, activation, nuclear translocation, and binding to the CYP2C11 promoter of the signal transducers mediating GH regulation of CYP2C11 expression, which dramatically contrasted with the complete unresponsiveness of the MSG-derived hepatocytes, also associated with hypermethylation of GH-response elements in the CYP2C11 promoter. Lastly, neonatal MSG treatment had no adverse effect on postnatal and adult testosterone levels. The results demonstrate that the sexually dimorphic expression of CYP2C11 is irreversibly imprinted shortly after birth by a hormone other than the customary testosterone, but likely by GH.

Technical and Anatomical Aspects of Retroperitoneoscopic Renal Surgery: A Summary of Tribulations and Resolutions Encountered at a Tertiary Care Institute of North India.

INTRODUCTION: Kidneys are a retroperitoneal organ but the widely practiced laparoscopic approach to renal surgery is transperitoneal due to the advantages of greater working space at the cost of entering the peritoneal cavity, risk of injury to intraperitoneal organs, and the increased risk of postoperative bowel complications. The classic open approach to kidney procedures has been the flank approach without violating the peritoneal cavity instead of the retroperitoneal approach to renal surgery with the advantages of direct access to the renal hilum, especially the renal artery. Being a technically challenging procedure, the retroperitoneoscopic approach is less practiced and needs an experienced surgical team. Through this study, we have tried to unveil the myths and illustrate the exact position of ports, which is the decisive initial step in retroperitoneoscopic surgery. MATERIAL AND METHODS: This retrospective study was conducted at a developing tertiary center in northern India with novice staff mainly to determine the technical and anatomical caveats pertaining to the retroperitoneoscopic approach for renal surgeries, the challenges faced, and their resolutions. The decision for the site of incision for primary or camera port was taken only after a proper anatomical study of the cadavers and ongoing retroperitoneal surgical experience while treating various patients suffering from renal diseases. The study comprised eight patients, during the period from June 2023 to March 2024. Various parameters, such as demographic variables, diagnosis, mean operative time, estimated blood loss, technical difficulties encountered and their resolution, complications, and reasons for conversion were studied. A total of 15 cadavers were dissected during the above time period to study finer anatomical details of port positioning and other details. RESULTS: After an elaborate study of 15 cadavers and thereafter performing surgery on eight patients during the above time period, surgery was successfully performed on six patients, and two patients needed conversion to open procedure due to dense adhesions and non-progression while complications occurred in two patients (peritoneal rent and renal vein injury), which were managed laparoscopically. CONCLUSION:  Nonetheless, restrictions of surgical space make retroperitoneoscopic space a challenging procedure but with elaborate experience, which we gained through cadaveric study, and surgical results obtained during the initial few cases such as the exact site of the primary port and technical intricacies, and handling of complications if and when faced, we hope our study will certainly make retroperitoneal space more amicable to urologists.

Protean Drainage Patterns of the Left Renal Vein: A Cadaveric and Retrospective Clinical Study on the Surgical Implications and Technical Feasibility.

BACKGROUND: The diverse drainage patterns of the left renal vein (LRV), often with asymptomatic congenital anomalies, present considerable challenges in renal and retroperitoneal surgical contexts. The potential for significant bleeding and subsequent renal compromise upon vascular injury highlights the need for increased surgical awareness. OBJECTIVE: This study investigates the LRV's variable anatomical drainage patterns and morphometry. It also evaluates the embryological factors contributing to these variations and discusses their surgical implications and technical considerations. METHODS: Anatomical dissections were conducted on 21 adult human cadavers within the Department of Anatomy. Concurrently, a retrospective analysis was conducted on 15 patients who underwent various retroperitoneal surgical interventions in the Urology Department. Demographic variables and intraoperative findings were recorded and analyzed. RESULTS: Dissection analysis predominantly identified preaortic LRVs in 18 cadavers. Notable anatomical variations included a circumaortic left renal vein (CLRV), a delayed preaortic confluence of extrahilar duo LRVs, and an extrahilar tetramerous confluence with a retroiliac topography. The majority of LRVs usually end in the inferior vena cava. However, an extrahilar tetramerous variant had an unusual drainage pathway. Out of 15 cases, three (20%) had a retroaortic left renal vein (RLRV). One patient with a nonfunctioning kidney had type 1 RLRV, and another patient with pelvic ureteric junction obstruction had type 4 retroiliac left renal vein (RILRV). In both of these patients, symptoms were relieved after surgery. In a young patient with left varicocele and microscopic hematuria who had type 2 RLRV, symptoms resolved spontaneously after a few months. CONCLUSION: A thorough understanding of the variable anatomical drainage patterns of the LRV is crucial for surgeons. Accurate preoperative identification can provide valuable insights, potentially leading to improved surgical outcomes in renal procedures.

Can periodontitis affect the COVID-19 severity, symptoms, hospital stay, and mortality? A case-control study.

Background/purpose: Poor oral hygiene and periodontal disease have been identified as potential risk factors for the coronavirus disease 2019 (COVID-19). The present study aimed to determine the association between periodontitis and COVID-19 severity, nature of symptoms, mortality, and hospital stay. Methods: In total, 163 COVID-19-positive patients (men: 93; women: 70) were categorized into two groups: the control group, consisting of 120 patients with asymptomatic or mild symptoms, and the case group, consisting of 43 patients with moderate-to-severe symptoms. The severity of periodontal disease, oral hygiene status (OHI), pocket depth (PD), bleeding on probing (BOP), number of decayed/missing/filled teeth, mortality, duration of stay in the hospital, oxygen requirement, and nature of COVID-19 symptoms were assessed in both groups. The association between periodontitis and COVID-19 was analyzed with other confounding factors such as age, sex, comorbidities, oral hygiene, and smoking status. Results: The presence of periodontitis increases the severity of COVID-19 by 3.7 times (p = 0.002). A statistically significant difference was noted for symptoms such as dizziness (p = 0.036), running nose/cold (p = 0.009), and headache (p = 0.005) in the presence of periodontitis. The risk estimate for death associated with periodontitis was 1.03. Additionally, the average duration of stay was longer for individuals with periodontitis than for those in the control group. Conclusion: There is a positive association between periodontal disease and COVID-19. Periodontitis increases the severity of COVID-19 and alters the symptoms. Hence, periodontal disease management should be an integral part of managing patients with coronavirus infection.

Dynamic Metal-Coordinated Adhesive and Self-Healable Antifreezing Hydrogels for Strain Sensing, Flexible Supercapacitors, and EMI Shielding Applications.

Dynamic metal-coordinated adhesive and self-healable hydrogel materials have garnered significant attention in recent years due to their potential applications in various fields. These hydrogels can form reversible metal-ligand bonds, resulting in a network structure that can be easily broken and reformed, leading to self-healing capabilities. In addition, these hydrogels possess excellent mechanical strength and flexibility, making them suitable for strain-sensing applications. In this work, we have developed a mechanically robust, highly stretchable, self-healing, and adhesive hydrogel by incorporating Ca2+-dicarboxylate dynamic metal-ligand cross-links in combination with low density chemical cross-links into a poly(acrylamide-co-maleic acid) copolymer structure. Utilizing the reversible nature of the Ca2+-dicarboxylate bond, the hydrogel exhibited a tensile strength of up to ∼250 kPa and was able to stretch to 15-16 times its original length. The hydrogel exhibited a high fracture energy of ∼1500 J m-2, similar to that of cartilage. Furthermore, the hydrogel showed good recovery, fatigue resistance, and fast self-healing properties due to the reversible Ca2+-dicarboxylate cross-links. The presence of Ca2+ resulted in a highly conductive hydrogel, which was utilized to design a flexible resistive strain sensor. This hydrogel can strongly adhere to different substrates, making it advantageous for applications in flexible electronic devices. When adhered to human body parts, the hydrogel can efficiently detect limb movements. The hydrogel also exhibited excellent performance as a solid electrolyte for flexible supercapacitors, with a capacitance of ∼260 F/g at 0.5 A/g current density. Due to its antifreezing and antidehydration properties, this hydrogel retains its flexibility at subzero temperatures for an extended period. Additionally, the porous network and high water content of the hydrogel impart remarkable electromagnetic attenuation properties, with a value of ∼38 dB in the 14.5-20.5 GHz frequency range, which is higher than any other hydrogel without conducting fillers. Overall, the hydrogel reported in this study exhibits diverse applications as a strain sensor, solid electrolyte for flexible supercapacitors, and efficient material for electromagnetic attenuation. Its multifunctional properties make it a promising candidate for use in various fields as a state-of-the-art material.

Diphenylmethyl selenocyanate attenuates malachite green induced oxidative injury through antioxidation & inhibition of DNA damage in mice.

BACKGROUND & OBJECTIVES: Malachite green (MG), an environmentally hazardous material, is used as a non permitted food colouring agent, especially in India. Selenium (Se) is an essential nutritional trace element required for animals and humans to guard against oxidative stress induced by xenobiotic compounds of diverse nature. In the present study, the role of the selenium compound diphenylmethyl selenocyanate (DMSE) was assessed on the oxidative stress (OS) induced by a food colouring agent, malachite green (MG) in vivo in mice. METHODS: Swiss albino mice (Mus musculus) were intraperitoneally injected with MG at a standardized dose of 100 µg/ mouse for 30 days. DMSE was given orally at an optimum dose of 3 mg/kg b.w. in pre (15 days) and concomitant treatment schedule throughout the experimental period. The parameters viz. ALT, AST, LPO, GSH, GST, SOD, CAT, GPx, TrxR, CA, MN, MI and DNA damage have been evaluated. RESULTS: The DMSE showed its potential to protect against MG induced hepatotoxicity by controlling the serum alanine aminotransferase and aspartate amino transferase (ALT and AST) levels and also ameliorated oxidative stress by modulating hepatic lipid peroxidation and different detoxifying and antioxidative enzymes such as glutathione-S-transferase (GST), superoxide dismutase (SOD), catalase (CAT), and also the selenoenzymes such as glutathione peroxidase (GPx) and thioredoxin reductase (TrxR) and reduced glutathione level which in turn reduced DNA damage. INTERPRETATION & CONCLUSIONS: The organo-selenium compound DMSE showed significant protection against MG induced heptotoxicity and DNA damage in murine model. Better protection was observed in pretreatment group than in the concomitant group. Further studies need to be done to understand the mechanism of action.

Extensive sex- and/or hormone-dependent expression of rat housekeeping genes.

AIM: Identify sex- and hormone-independent housekeeping genes in rat liver by using a commercially available quantitative reverse transcription-polymerase chain reaction array designed to measure the expression of 32 rat housekeeping genes. RESULTS: We found that the levels of five of the genes were sexually dimorphic, 22 genes were overexpressed, and one was underexpressed in multi-hormone-deficient hypophysectomized rats of both sexes. Only three genes fulfilled the stability criteria determined by geNorm and NormFinder as suitable housekeeping genes. Normalizing quantitative reverse transcription-polymerase chain reaction data with either of these three genes alone, the geometric means of any two of the genes, or even the geometric mean of all the three genes, produced similar results. In contrast, application of unproven housekeeping genes could lead to erroneous conclusions, having found that insulin-like growth factor 1 messenger RNA levels could be calculated dramatically either as male or as female predominant depending on the choice of housekeeping gene. CONCLUSION: It is essential to validate the constancy of housekeeping genes under every experimental condition. (This research protocol was approved by the university's Institutional Animal Care and Use Committee.).

Synthesis of [1,3, NH2-(15)N3] (5'S)-8,5'-cyclo-2'-deoxyguanosine.

To facilitate NMR studies and low-level detection in biological samples by mass spectrometry, [1,3, NH2-(15)N3] (5'S)-8,5'-cyclo-2'-deoxyguanosine was synthesized from imidazole-4,5-dicarboxylic acid in 21 steps. The three (15)N isotopes were introduced during the chemo-enzymatic preparation of [1,3, NH2-(15)N3]-2'-deoxyguanosine using an established procedure. The (15)N-labeled 2'-deoxyguanosine was converted to a 5'-phenylthio derivative, which allowed the 8-5' covalent bond formation via photochemical homolytic cleavage of the C-SPh bond. SeO2 oxidation of C-5' followed by sodium borohydride reduction and deprotection gave the desired product in good yield. The isotopic purity of the [1,3, NH2-(15)N3] (5'S)-8,5'-cyclo-2'-deoxyguanosine was in excess of 99.94 atom% based on liquid chromatography-mass spectrometry measurements.

Bilateral Symmetric Absence of Lumbricals of the Foot: A Rare Case Report.

Variations of the lumbricals of the foot are not common, unlike the lumbricals of the hand. Few cases of unilateral absence of lumbricals of the foot have been reported. There have been far fewer cases of bilateral asymmetric absence of the lumbricals of the foot reported. The study reports the findings observed during the routine dissection for undergraduate medical students in the Department of Anatomy. We observed that the third lumbrical in both the feet was absent in an elderly male cadaver. The other features of both feet were usual. Externally, there was no apparent deformity, operation mark, or evidence of injury in any of the feet. Considering the functional role of the lumbricals in particular and other small intrinsic muscles in general, on the mechanics of foot movement, it is of much significance to have the knowledge of the possible variations and their clinical implications, which should be studied using different diagnostic tools such as imaging techniques, dynamometry, and other modalities.

Structures of (5'S)-8,5'-Cyclo-2'-deoxyguanosine Mismatched with dA or dT.

Diastereomeric 8,5'-cyclopurine 2'-deoxynucleosides, containing a covalent bond between the deoxyribose and the purine base, are induced in DNA by ionizing radiation. They are suspected to play a role in the etiology of neurodegeneration in xeroderma pigmentosum patients. If not repaired, the S-8,5'-cyclo-2'-deoxyguanosine lesion (S-cdG) induces Pol V-dependent mutations at a frequency of 34% in Escherichia coli. Most are S-cdG → A transitions, suggesting mis-incorporation of dTTP opposite the lesion during replication bypass, although low levels of S-cdG → T transversions, arising from mis-incorporation of dATP, are also observed. We report the structures of 5'-d(GTGCXTGTTTGT)-3'·5'-d(ACAAACAYGCAC)-3', where X denotes S-cdG and Y denotes either dA or dT, corresponding to the situation following mis-insertion of either dTTP or dATP opposite the S-cdG lesion. The S-cdG·dT mismatch pair adopts a wobble base pairing. This provides a plausible rationale for the S-cdG → A transitions. The S-cdG·dA mismatch pair differs in conformation from the dG·dA mismatch pair. For the S-cdG·dA mismatch pair, both S-cdG and dA intercalate, but no hydrogen bonding is observed between S-cdG and dA. This is consistent with the lower levels of S-cdG → T transitions in E. coli.

Stability of N-glycosidic bond of (5'S)-8,5'-cyclo-2'-deoxyguanosine.

8,5'-Cyclopurine deoxynucleosides are unique tandem lesions containing an additional covalent bond between the base and the sugar. These mutagenic and genotoxic lesions are repaired only by nucleotide excision repair. The N-glycosidic (or C1'-N9) bond of 2'-deoxyguanosine (dG) derivatives is usually susceptible to acid hydrolysis, but even after cleavage of this bond of the cyclopurine lesions, the base would remain attached to the sugar. Here, the stability of the N-glycosidic bond and the products formed by formic acid hydrolysis of (5'S)-8,5'-cyclo-2'-deoxyguanosine (S-cdG) were investigated. For comparison, the stability of the N-glycosidic bond of 8,5'-cyclo-2',5'-dideoxyguanosine (ddcdG), 8-methyl-2'-deoxyguanosine (8-Me-dG), 7,8-dihydro-8-oxo-2'-deoxyguanosine (8-Oxo-dG), and dG was also studied. In various acid conditions, S-cdG and ddcdG exhibited similar stability to hydrolysis. Likewise, 8-Me-dG and dG showed comparable stability, but the half-lives of the cyclic dG lesions were at least 5-fold higher than those of dG or 8-Me-dG. NMR studies were carried out to investigate the products formed after the cleavage of the C1'-N9 bond. 2-Deoxyribose generated α and ß anomers of deoxyribopyranose and deoxyribopyranose oligomers following acid treatment. S-cdG gave α- and ß-deoxyribopyranose linked guanine as the major products, but α and ß anomers of deoxyribofuranose linked guanine and other products were also detected. The N-glycosidic bond of 8-Oxo-dG was found exceptionally stable in acid. Computational studies determined that both the protonation of the N7 atom and the rate constant in the bond breaking step control the overall kinetics of hydrolysis, but both varied for the molecules studied indicating a delicate balance between the two steps. Nevertheless, the computational approach successfully predicted the trend observed experimentally. For 8-Oxo-dG, the low pK(a) of O(8) and N3 prevented appreciable protonation, making the free energy for N-glycosidic bond cleavage in the subsequent step very high.

Procedural sedation in children for fractionated radiation treatment: Intranasal dexmedetomidine versus oral midazolam and ketamine.

Background and Aims: Sedation in paediatric cancer for fractionated radiation treatment (RT) is unique as the child has to be still for accurate delivery of RT, monitoring of the child is from a remote location and sedation is repeated for multiple sessions of RT. The present study was undertaken to compare the efficacy of intranasal dexmedetomidine with oral midazolam and ketamine combination for repeated sedation during fractionated RT in paediatric oncology. Methods: Ninety children aged between 3-6 years, planned for 21 fractions of RT, were randomised to receive intranasal dexmedetomidine 2 µg/kg (group D) or oral midazolam 0.2 mg/kg and ketamine 5 mg/kg (group MK). The 21 sessions of fractionated radiotherapy were divided into three subgroups of seven consecutive exposures 1-7, 8-14 and 15-21 for comparison. The primary endpoint was to determine the incidence of successful sedation. The sedation score achieved, time to satisfactory sedation and discharge, rescue ketamine required, and side effects were secondary endpoints. Results: The incidence of successful sedation in the three successive RT subgroups; sessions: 1-7, 8-14 and 15-21, was 82%, 75.6% and 66.7% in group D, as compared to 40%, 24.4% and 13.3% in group MK, respectively. (P < 0.001). A decrease in successful sedation was noted in the successive subgroups. Time to successful sedation and discharge was earlier in group D in comparison to MK (P = 0.000). More patients in group MK required rescue ketamine (P = 000). Conclusion: Intranasal dexmedetomidine produces more satisfactory sedation as compared to oral ketamine with midazolam for fractionated RT.

Bilateral higher carotid termination with rare anomalous emergence of ventral branches of the external carotid arteries: A cadaver study.

The common carotid artery bifurcates into two terminal branches-the external and internal carotid arteries. The head, neck, and face regions principally get their blood supply from the external carotid artery and its branches. Some previous articles have mentioned the abnormal pattern of the external carotid artery branching and its variable origin. In this article, a rare case has been documented, having the combination of anomalies of the high carotid termination and bilateral variable origin of the ventral branches of the external carotid artery encountered during routine dissection of the head and neck region of a 55-year-old male cadaver in the Department of Anatomy. In this instance, on the right side of the neck, we observed the presence of a thyrolinguofacial trunk which arose from the ventral surface of the external carotid artery, and on the left side, the linguofacial trunk emerged from the ventral surface of the external carotid artery while the superior thyroid artery branched off directly from the left common carotid artery. This apart, there was bilateral high termination of the common carotid artery. Although the exact embryogenesis of such common arterial trunks anomalies and high carotid termination is not clear, detailed and precise anatomical knowledge of such a combination of anomalies will provide further insight for better radiological evaluation and to avert iatrogenic vascular injuries during any surgical procedures of the head and face region.

Bilateral High Trifurcation of the Common Carotid Artery and Variable Emergence of the Lower Branches of the External Carotid Artery: A Cadaveric Case Report.

Trifurcation of the common carotid artery in the neck region is a rare anatomical variation. In the present study, we reported a rare case having the combination of anomalies of the bilateral high common carotid arteries trifurcation and variable origin of lower branches of the external carotid artery during routine dissection of the head and neck region of a 60-year-old male cadaver in the Department of Anatomy. Both on the left and right sides of the neck region, the common carotid artery gave off three terminal branches: internal carotid artery, external carotid artery, and ascending pharyngeal arteries. Further, we also observed the presence of bilateral linguofacial trunks (common arterial trunks) that emerged from the external carotid arteries and also the left superior thyroid artery that originated directly from the left common carotid artery. Even though the embryogenesis of the variable origin of such arterial trunks is not apparent, it is very indispensable to have sound knowledge and better comprehension of the accurate anatomical architecture of such a rare combination of carotid arterial system anomalies for correct interpretation of the vascular imaging that pave the pathway for successful execution of surgical interventions in the neck region because of its utmost clinical implication.

(5'S)-8,5'-cyclo-2'-deoxyguanosine is a strong block to replication, a potent pol V-dependent mutagenic lesion, and is inefficiently repaired in Escherichia coli.

8,5'-Cyclopurines, making up an important class of ionizing radiation-induced tandem DNA damage, are repaired only by nucleotide excision repair (NER). They accumulate in NER-impaired cells, as in Cockayne syndrome group B and certain Xeroderma Pigmentosum patients. A plasmid containing (5'S)-8,5'-cyclo-2'-deoxyguanosine (S-cdG) was replicated in Escherichia coli with specific DNA polymerase knockouts. Viability was <1% in the wild-type strain, which increased to 5.5% with SOS. Viability decreased further in a pol II(-) strain, whereas it increased considerably in a pol IV(-) strain. Remarkably, no progeny was recovered from a pol V(-) strain, indicating that pol V is absolutely required for bypassing S-cdG. Progeny analyses indicated that S-cdG is significantly mutagenic, inducing ~34% mutation with SOS. Most mutations were S-cdG → A mutations, though S-cdG → T mutation and deletion of 5'C also occurred. Incisions of purified UvrABC nuclease on S-cdG, S-cdA, and C8-dG-AP on a duplex 51-mer showed that the incision rates are C8-dG-AP > S-cdA > S-cdG. In summary, S-cdG is a major block to DNA replication, highly mutagenic, and repaired slowly in E. coli.

Structure of (5'S)-8,5'-cyclo-2'-deoxyguanosine in DNA.

Diastereomeric 8,5'-cyclopurine 2'-deoxynucleosides, containing a covalent bond between the deoxyribose and the purine base, represent an important class of DNA damage induced by ionizing radiation. The 8,5'-cyclo-2'-deoxyguanosine lesion (cdG) has been recently reported to be a strong block of replication and highly mutagenic in Escherichia coli. The 8,5'-cyclopurine-2'-deoxyriboses are suspected to play a role in the etiology of neurodegeneration in xeroderma pigmentosum patients. These lesions cannot be repaired by base excision repair, but they are substrates for nucleotide excision repair. The structure of an oligodeoxynucleotide duplex containing a site-specific S-cdG lesion placed opposite dC in the complementary strand was obtained by molecular dynamics calculations restrained by distance and dihedral angle restraints obtained from NMR spectroscopy. The S-cdG deoxyribose exhibited the O4'-exo (west) pseudorotation. Significant perturbations were observed for the ß, γ, and χ torsion angles of the S-cdG nucleoside. Watson-Crick base pairing was conserved at the S-cdG·dC pair. However, the O4'-exo pseudorotation of the S-cdG deoxyribose perturbed the helical twist and base pair stacking at the lesion site and the 5'-neighbor dC·dG base pair. Thermodynamic destabilization of the duplex measured by UV melting experiments correlated with base stacking and structural perturbations involving the modified S-cdG·dC and 3'- neighbor dT·dA base pairs. These perturbations may be responsible for both the genotoxicity of this lesion and its ability to be recognized by nucleotide excision repair.