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BackgroundTo document the role of sub-clinical infections in disc disorders and investigate the existence of microbiome in intervertebral discs (IVD).MethodsGenomic DNA from 24 lumbar IVDs [8—MRI normal discs (ND) from brain dead yet alive organ donors, 8—disc herniation (DH), 8—disc degeneration (DD)] was subjected to 16SrRNA sequencing for profiling the diversity of human disc microbiome in health and disease. The disc microbiome was further compared to established human gut and skin microbiomes.ResultsAll healthy MRI normal discs from brain dead yet alive organ donors also had a rich bacterial presence. A total of 424 different species (355-ND, 346-DD, and 322-DH) were detected, with 42.75% OTUs being classified at kingdom level, 44% at the phylum level, 22.62% at genus level, and 5.5% at species level. Varying biodiversity and abundance between healthy and diseased discs were documented with protective bacteria being abundant in normal discs, and putative pathogens abundant in DD and DH. Propionibacterium acnes had a similar but lower abundance to other pathogens in all three groups ND (3.07%), DD (3.88%), DH (1.56%). Fifty-eight bacteria were common between gut and IVD microbiomes, 29 between skin and IVD microbiomes, and six common to gut/skin/IVD.ConclusionOur study challenges the hitherto concept of sterility in healthy IVD and documented a microbiome even in MRI normal healthy discs. The varying abundance of bacteria between ND, DD, and DH documents ‘dysbiosis’ as a possible etiology of DD. Many known pathogens were identified in greater abundance than Propionibacterium acnes, and there was evidence for the presence of the gut/skin/spine microbiome axis.

Kappaphycus alvarezii seaweed extract (KSWE) is known to enhance crop productivity and impart stress tolerance. Close to one quarter of foliar spray applied to maize falls on the soil, either as drift or from leaf as drip. It was hypothesized that the drift spray would profoundly influence soil microbes under stress. An experiment was conducted with five treatments, with or without KSWE application at critical stages of maize grown under soil moisture stress and compared with an irrigated control. An Illumina platform was employed for the analysis of the V3–V4 region of 16S rRNA gene from the soil metagenome. A total of 345,552 operational taxonomic units were generated which were classified into 55 phyla, 152 classes, 240 orders, 305 families and 593 genera. Shannon’s index and Shannon’s equitability indicated increased soil bacterial diversity after multiple KSWE applications under conditions of abiotic duress. The abundance of the genera Alicyclobacillus, Anaerolinea, Bacillus, Balneimonas, Nitrospira, Rubrobacter and Steroidobacter decreased (49–79%) under drought imposed at the V5,10 and 15 stages of maize over the irrigated control, while it significantly improved when followed by KSWE application under drought. Flavobacterium, Nitrosomonas, Nitrosovibrio, Rubrobacter genera and several other bacterial taxa which are important for plant growth promotion and nutrient cycling were found to be enriched by KSWE application under drought conditions. Treatments having enriched microbial abundance due to KSWE application under stress recorded higher soil enzymatic activities and plant cob yield, suggesting the contribution of altered soil ecology mediated by KSWE as one of the reasons for improvement of yield.

PurposeTo probe the pathophysiological basis of Modic change (MC) by multimodal imaging rather than by MRI alone.MethodsNineteen radiological signs found in mild infections and traumatic endplate fractures were identified by MRI and CT, and by elimination, three signs unique to infection and trauma were distilled. By ranking the Z score, radiological ‘Endplate Infection Probability Score’ (EIPS) was developed. The score’s ability to differentiate infection and traumatic endplate changes (EPC) was validated in a fresh set of 15 patients each, with documented infection and trauma. The EIPS, ESR, CRP, and Numeric Pain Rating Scale (NRS) were then compared between 115 patients with and 80 patients without MC.ResultsThe EIPS had a confidence of 66.4%, 83% and, 100% for scores of 4, 5 and, 6, respectively, for end plate changes suggesting infection. The mean EIPS was 4.85 ± 1.94 in patients with Modic changes compared to − 0.66 ± 0.49 in patients without Modic changes (p < 0.001). Seventy-eight (67.64%) patients with MC had a score of 6, indicating high infection possibility. There was a difference in the NRS (p < 0.01), ESR (p = 0.05), CRP (p < 0.01), and type of pain (p < 0.01) between patients with and without MC.ConclusionMultimodal imaging showed many radiological signs not easily seen in MRI alone and thus missed in Modic classification. There were distinct radiological differences between EPCs of trauma and infection which allowed the development of an EIPS. The scores showed that 67.64% of our study patients with Modic changes had EPCs resembling infection rather than trauma suggesting the possibility of an infective aetiology and allowing us to propose an alternate theory of ‘Primary Endplatitis’.

Degeneration of the intervertebral disc is associated with a decrease in extra-cellular matrix (ECM) content due to an imbalance in anabolic and catabolic signaling. Our previous study profiled the core matrisome of fetal NP’s and identified various proteins with anabolic potential for regenerative therapies. This study aims to complement those results by exploring ECM regulators, associated proteins and secreted factors of the fetal nucleus pulposus (NP). Proteomic data of 9 fetal, 7 healthy adults (age 22–79), and 11 degenerated NP’s was analyzed. Based on the selection criteria, a total of 45 proteins were identified, of which 14 were uniquely expressed or upregulated in fetus compared to adult NP’s. Pathway analysis with these proteins revealed a significant upregulation of one pathway and two biological processes, in which 12 proteins were involved. Prolyl 4 hydroxylase (P4HA) 1 and 2, Procollagen-lysine, 2-oxoglutarate 5-dioxygenase (PLOD) 1, and Heat shock protein 47 (SERPINH1) were involved in ‘collagen biosynthesis’ pathway. In addition, PLOD 1, SERPINH1, Annexin A1 and A4, CD109 and Galectin 3 (LGALS3) were all involved in biological process of ‘tissue development’. Furthermore Annexin A1, A4 and A5, LGALS-3 and SERPINF1 were featured in ‘negative regulation of cell death’. In conclusion, additionally to core ECM proteome, this study reveals ECM regulators and ECM affiliated proteins of interest to study for regenerative therapies, and their potential should be validated in future mechanistic experiments.

Seaweed extracts have been reported to be effective crop biostimulants having low carbon foot print. However, the mechanism of their action on crops has remained elusive due to dearth of studies. Kappaphycus alvarezii seaweed extract (K-sap) has been found to increase the yield of several crops as well as impart drought tolerance. Here, an attempt was made to get a global view of the transcriptome response of maize roots when subjected to soil drench application of K-sap under normally irrigated and drought conditions. mRNA were sequenced using high-throughput RNA sequencing employing Illumina platform and transcriptome mapping was carried out. K-sap applied plants under drought, when compared to its control, recorded several differentially expressed genes involved in RNA-, protein-, cell wall–, signaling-, transport-, stress-, development-, cell-, secondary metabolism–, hormone metabolism–, DNA-, lipid metabolism–, major and minor CHO metabolism–, redox-, metal handling–, amino acid metabolism–, nucleotide metabolism–, TCA-, and N metabolism–related pathways. Up-regulation was observed in the genes coding towards enhancement of root growth, gibberellic acid and auxin signaling, seed development, nitrogen metabolism, transport, and antioxidant activity like glutathione S-transferase and peroxidases. On the other hand, down-regulation of starch and sucrose degradation–related transcripts was apparent. These related well to our morphological observations on improved root growth, grain yield, and higher nutrient content of the roots under drought stress owing to application of K-sap. The study gives a comprehensive insight of several biological processes being modulated and opens up avenues for further targeted mechanistic studies.

PurposeTo quantitatively assess the dynamic changes of Lactate in lumbar discs under different physiological conditions using MRS and T2r.MethodsIn step1, MRS and T2r sequences were standardized in 10 volunteers. Step2, analysed effects of high cellular demand. 66 discs of 20 volunteers with no back pain were evaluated pre-exercise (EX-0), immediately after targeted short-time low back exercises (EX-1) and 60 min after (EX-2). In Step 3, to study effects of high glucose and oxygen concentration, 50 lumbar discs in 10 volunteers were analysed before (D0) and after 10 days intake of the calcium channel blocker, nimodipine (D1).ResultsLactate showed a distinctly different response to exercise in that Grade 1 discs with a significant decrease in EX-1 and a trend for normalization in Ex-2. In contrast, Pfirrmann grade 2 and 3 and discs above 40 years showed a higher lactate relative to proteoglycan in EX-0, an increase in lactate EX-1 and mild dip in Ex-2. Similarly, following nimodipine, grade 1 discs showed an increase in lactate which was absent in grade 2 and 3 discs. In contrast, exercise and Nimodipine had no significant change in T2r values and MRS spectrum of proteoglycan, N-acetyl aspartate, carbohydrate, choline, creatine, and glutathione across age groups and Pfirrmann grades.ConclusionMRS documented changes in lactate response to cellular demand which suggested a ‘Lactate Symbiotic metabolic Pathway’. The differences in lactate response preceded changes in Proteoglycan/hydration and thus could be a dynamic radiological biomarker of early degeneration.

Transnasal endoscopic cranial base surgery is a novel minimal-access method for reaching the midline cranial base. Postoperative cerebrospinal fluid leak remains a persistent challenge. A new method for watertight closure of the anterior cranial base is presented. To achieve watertight closure of the anterior cranial base, autologous fascia lata was used to create a \"gasket seal\" around a bone buttress, followed by application of a tissue sealant such as DuraSeal (Confluent Surgical, Inc., Waltham, MA). The gasket-seal closure was used to seal the anterior cranial base in a series of 10 patients with intradural surgery for suprasellar craniopharyngiomas (n = 5), planum meningiomas (n = 3), clival chordoma (n = 1), and recurrent iatrogenic cerebrospinal fluid leak (n = 1). Lumbar drains were placed intraoperatively in five patients and remained in place for 3 days postoperatively. After a mean follow-up period of 12 months, there were no cerebrospinal fluid leaks. The gasket-seal closure is an effective method for achieving watertight closure of the anterior cranial base after endoscopic intradural surgery.

Kappaphycus alvarezii seaweed extract (KSWE) has been known for its plant biostimulant and stress alleviation activities on various crops. However, very few reports are available depicting its impact at the molecular level, which is crucial in identifying the mechanism of action of KSWE on plants. Here, maize leaf tissue of control and KSWE-treated plants were analyzed for their transcriptional changes under drought stress. KSWE was applied foliarly at the V5 stage of maize crop under drought, and leaf transcriptome analysis was performed. It was found that a total of 380 and 631 genes were up- and downregulated, respectively, due to the application of KSWE. Genes involved in nitrate transportation, signal transmission, photosynthesis, transmembrane transport of various ions, glycogen, and starch biosynthetic processes were found upregulated in KSWE-treated plants, while genes involved in the catabolism of polysaccharide molecules such as starch as well as cell wall macromolecules like chitin and protein degradation were found downregulated. An overview of differentially expressed genes involved in metabolic as well as regulatory processes in KSWE-treated plants was also analyzed via Mapman tool. Phytohormone signaling genes such as cytokinin-independent 1 (involved in cytokine signal transduction), Ent-kaurene synthase and GA20 oxidase (involved in gibberellin synthesis), and gene of 2-oxoglutarate-dependent dioxygenase enzyme activity (involved in ethylene synthesis) were found upregulated while 9-cis-epoxycarotenoid dioxygenase (a gene involved in abscisic acid synthesis) was found downregulated due to the application of KSWE. Modulation of gene expression in maize leaf tissue in response to KSWE treatment elucidates mechanisms to ward off drought stress, which can be extended to understand similar phenomenon in other crops as well. This molecular knowledge can be utilized to make the use of KSWE more efficient and sustainable.

A pot experiment was conducted in a split-plot design with five concentrations (0, 5, 10, 15, and 20%) of Kappaphycus alvarezii seaweed extract (KSWE) as main plots and two black gram varieties (TAU–1 and DBGV–5) as subplots, with treatment each replicated thrice. Activities of superoxide dismutase (SOD), catalase (CAT), glutathione reductase (GR), and ascorbate peroxidase (APX), protein, non-protein and total thiol contents in black gram root nodules were evaluated at 23–24 days after sowing (DAS), 72–96 h after a single foliar application of the extract at 20 DAS. SOD and GR activities were consistently higher in DBGV–5 than in TAU–1, with SOD declining at elevated KSWE concentrations and GR showing a pronounced reduction at 15% KSWE in 2020. APX activity was reduced by KSWE in 2019 but remained unaffected in 2020, whereas CAT activity was stable across treatments and years. Overall, KSWE modulated nodule antioxidant enzyme responses in a concentration-, variety-, and year-dependent manner, with DBGV–5 exhibiting greater enzymatic sensitivity. Thiol metabolism was likewise influenced by KSWE and genotype, with significant increases in total and protein thiols, particularly at 5% KSWE, and consistently higher levels in DBGV–5 than in TAU–1. Non-protein thiols showed significant variety × dose interactions in 2019 but remained unchanged in 2020, indicating a context-dependent adjustment of cellular redox status. The present study represents one of the earliest evaluations of the effects of seaweed biostimulants on antioxidant responses in root nodules.

Study Design Observational comparative study. Objective To study the role of magnetic resonance spectroscopy (MRS) and T2 relaxometry (T2r) as imaging biomarkers for identifying early lumbar disc degeneration. Methods We evaluated 236 discs in normal volunteers and 215 discs in low back pain (LBP) patients by MRS and T2r to document the molecular spectra of various metabolites as well as disc hydration and collagen content, respectively. All volunteer discs were Pfirrmann grade 1 (PF1), whereas patients with LBP had PF 1 (n = 156) and PF 2 (n = 59). The study population was compared in three age groups: A (20-30 years), B (30-40 years), and C (40-50 years). Results T2r, an indicator of collagen and hydration, was higher in volunteers (121.8 ± 31.1), compared to PF 1 patients (110.68 ± 23.96) and PF 2 patients (90.15 ± 25.81) (P = 0.001). Proteoglycan assessed by MRS was more stable for volunteers (3.39 ± 1.69) and PF 1 patients (3.6 ± 1.69) but reduced in PF 2 patients (2.86 ± 1.47), showing that structural molecules did not alter within the PF 1. However, lactate and other metabolites showed a difference even within PF1 between volunteers and LBP patients. We were able to identify a unique subset of PF 1 that had a normal value of proteoglycan and T2r but altered metabolite distribution, which may represent early disc degeneration (DD). Conclusion MRS and T2r can be used as imaging biomarkers for early DD by identifying altered metabolic activity with an intact matrix.