Role of Serum Micro-RNA-122-5p Expression as a Circulatory Biomarker in People Having Both Knee Osteoarthritis and Osteoporosis: A Case-Control Study.

Background Although knee osteoarthritis (KOA) and osteoporosis (OP) manifest distinct pathophysiologies, they share numerous similarities. These health conditions are commonly found in older individuals, particularly among women. The objective of this study is to explore the expression of micro-RNA (miRNA) 122-5p (miR-122-5p) in people affected by both KOA and OP. The main aim is to identify diagnostic biomarkers and potential therapeutic targets, which could help develop personalized treatment approaches. Methods As part of the study, a total of 268 serum samples were collected from the participants, who were divided into four groups: KOA, OP, KOA and OP, and controls, with 67 subjects per group. The miRNA species-containing total RNA was isolated from the serum samples using an miRNeasy serum/plasma kit by QIAGEN (Hilden, Germany). The expression of miR-122-5p was examined in each group using real-time quantitative polymerase chain reaction. Results Expression of miR-122-5p in all three groups (KOA, OP, and common group of KOA and OP) was significantly upregulated, and the fold change value was much higher in the group having both diseases. Conclusions These results might contribute to the identification of cases at risk, early diagnosis, and development, and might also contribute to the development of therapeutic targets in subjects having both KOA and OP.

Prevalence of Abnormal Bone-Specific Alkaline Phosphatase in Orthopaedic Trauma Patients: A Cross-Sectional Study From a Tertiary Trauma Centre.

INTRODUCTION: Bone-specific alkaline phosphatase (BALP) has been a sensitive and reliable marker of bone metabolic activity. The study aimed to study the epidemiology of orthopaedic trauma patients and estimate the level of BALP in these patients. MATERIAL AND METHODS: This hospital-based observational cross-sectional study was conducted on over 300 patients admitted to the orthopaedic trauma unit in the level I trauma centre of North India during the study period from January 2020 to January 2021. The level of BALP was assessed in serum samples of 258 patients. RESULT: A total of 300 patients were included in this study, with an average age of 33.52±17.16 years. Road traffic accidents were the most common mode of injury among admitted patients. We found a significant linear correlation between age and mean BALP levels (P-value < 0.005). The mean age of patients having abnormal BALP is 63.0±16.61 years. Females (22.61±16.23) had greater mean BALP levels than males (13.15±5.86). Among different fractures, fractures around the hip had a higher mean BALP level (23.17±15.07). Patients with diabetes mellitus have higher mean BALP levels. CONCLUSION: BALP is a cost-effective, readily available bone marker to assess the metabolic activity of orthopaedic trauma patients. Old age patients should undergo routine BALP estimation at admission to rule out metabolic disorders, and timely intervention will prevent complications associated with metabolic disorders.

Expression of Antimicrobial Peptides and Cytokines in Human Omentum Following Abdominal Surgery.

Introduction Omentum can secrete out biological agents like different growth factors, cytokines, and antimicrobial peptides. The aim of our study was to determine the expression of antimicrobial peptides and cytokines in human omentum tissue and its response to intra-abdominal infection. Methodology Omentum tissue was obtained from 60 patients: control (n=20) and cases (n=40). mRNA expression of antimicrobial peptides (LL-37, HBD-1, HBD-2, HNP1-3) and cytokines (TNF- α, IL-8, IL-10, IL1ß) was evaluated using Real-Time PCR. Protein quantification was done by Immunoblotting and ELISA. Results Significantly higher expression of antimicrobial peptides (LL-37, HBD-1, HBD-2, HNP1-3) and cytokines (TNF- α, IL-8, IL-10, IL1ß) was observed in cases as compared to control at both the transcriptional and translational level (p<0.0001). Conclusion Omentum governs a population of antimicrobial peptides with potent immunologic functions. The expression of antimicrobial peptides and cytokines is inducible and increases with the severity of infection. Omentum is thus an immunologically active and adaptable organ but its complete regulatory mechanism is still elusive.

Prevalence of Vitamin D Deficiency in a Young Adult With Acute Spinal Cord Injury.

INTRODUCTION: Vitamin D deficiency has been implicated as an etiologic factor responsible for osteoporosis and various skeletal and extra-skeletal issues in spinal cord injury patients. There is a dearth of publications regarding the prevalence of vitamin D deficiency in acute spinal cord injury (ASCI) patients, thus it becomes imperative to study the status of vitamin D in ASCI cases to make an early diagnosis and start treatment for osteoporosis. Apart from this, we also planned to evaluate other factors associated with vitamin D deficiency in our subset of patients. MATERIAL AND METHODS:  This cross-sectional cohort study included patients with acute thoracolumbar spinal cord injury patients admitted to a tertiary trauma centre between July 2019 and July 2020. Patients were assessed clinically and classified as per the American Spinal Cord Injury Association (ASIA) scale. Demographic details along with the mode of trauma and duration of injury were noted. Serum 25(OH) vitamin D3 levels were measured by chemiluminescence immunoassay. Depending upon serum 25(OH) vitamin D3 level, patients were classified into vitamin D deficient with serum level less than 20 ng/ml, vitamin D insufficient with serum level between 21-29 ng/ml and vitamin D sufficient with serum level greater than 30 ng/ml. RESULTS: Mean vitamin D level in 85 ASCI subjects (mean age 30.82 ± 6.77 years, 60 males) was 20.56 ± 11.22 ng/ml. Fifty subjects (58.82%) were vitamin D deficient, 15 subjects (17.64%) were vitamin D insufficient and the rest (n=20, 23.52%) were vitamin D sufficient. There was no significant difference in vitamin D levels as per gender, age, mode of trauma, type of injury and injury location. Patients admitted on the fifth day of injury had maximum vitamin D levels (mean 25.7143 ± 8.32 ng/ml), but it was also insignificant. The mean vitamin D level of subjects with samples taken during the summer season was significantly higher as compared to the winter season (p value <.05). CONCLUSION: Vitamin D deficiency is widely prevalent in ASCI patients at admission to the trauma centre. Seventy-six percent of patients had vitamin D levels below 30 ng/ml in our study. Routine measurement of 25(OH) vitamin D3 levels at the time of admission is recommended for early diagnosis of vitamin D deficiency. Early treatment will be helpful in the prevention of osteoporosis and its long-term related consequences.

nNOS induction and NOSIP interaction impact granulopoiesis and neutrophil differentiation by modulating nitric oxide generation.

Nitric oxide (NO), a versatile free radical and a signalling molecule, plays an important role in the haematopoiesis, inflammation and infection. Impaired proliferation and differentiation of myeloid cells lead to malignancies and Hematopoietic deficiencies. This study was aimed to define the role of nNOS derived NO in neutrophil differentiation (in-vitro) and granulopoiesis (in-vivo) using multipronged approaches. The results obtained from nNOS over-expressing K562 cells revealed induction in C/EBPα derived neutrophil differentiation as evident by an increase in the expression of neutrophil specific cell surface markers, genes, transcription factors and functionality. nNOS mediated response also involved G-CSFR-STAT-3 axis during differentiation. Consistent increase in NO generation was observed during neutrophil differentiation of mice and human CD34+ HSPCs. Furthermore, granulopoiesis was abrogated in the nNOS inhibitor treated mice, depicting a decrease in the numbers of BM mature and progenitor neutrophils. Likewise, in vitro inhibition of nNOS in human CD34+ HSPCs indicated an indispensable role of nNOS in neutrophil differentiation. Expression of nNOS inhibitory protein, NOSIP was significantly and consistently decreased during the final stage of differentiation and was linked with the augmentation in NO release. Moreover, neutrophils from CML patients had more NOSIP and less NO generation as compared to the PMNs from healthy individuals. The present study thus indicates a critical role of nNOS, and its interaction with NOSIP during neutrophil differentiation. The study also highlights the importance of nNOS in the neutrophil progenitor proliferation and differentiation warranting investigations to assess its role in the haematopoiesis-related disorders.

Calcitriol attenuates TLR2/IL-33 signaling pathway to repress Th9 cell differentiation and potentially limits the pathophysiology of rheumatoid arthritis.

There is limited information regarding the TLR2 signaling pathway involved in Th9 cell differentiation. The role of calcitriol in regulating TLR2-mediated Th9 cell development is unknown. Thus, we aimed to unravel the TLR2 signaling pathway in Th9 cells and its regulation by calcitriol. We have used n = 5-6 animals for each murine experiment. Human studies involved five healthy volunteers. Moreover, ten healthy individuals and ten RA patients were included in the study. Murine and human Th9 cells were treated with Calcitriol (100 nM) and Pam3CSK4 (2 µg/mL). The number of IL-9+ve cells was determined by flow cytometry. Real-time PCR was used to assess the gene expression. Serum 25(OH)D3 levels were determined by HPLC. We observed that TLR2 signals via IL-33/ST2 in Th9 cells. Increased TLR2 expression associated with increased IL9 expression and augmented disease severity in RA patients. Calcitriol attenuated TLR2 signaling in murine and human Th9 cells. Low serum vitamin D3 level negatively associated with increased IL-9 and TLR2 expression and disease severity in RA patients. Our data suggest a potential role of calcitriol to ameliorate the disease severity of RA patients.

Management of Grade IV Pressure Ulcers With a Novel Negative Pressure Device in Traumatic Paraplegia Subjects.

Introduction Pressure ulcers (PUs) are a major health problem for bedridden patients or persons with reduced mobility; individuals with spinal cord injury (SCI) are more prone to developing pressure ulcers. The purpose of this study was to determine the efficacy of a novel negative pressure wound therapy (NPWT) system for the treatment of Grade IV PUs. Methods A total of 34 SCI patients with Grade IV PUs were divided into two groups: 17 cases were managed by our bellows-powered negative pressure device (NPD) and 17 received wet-to-moist gauze dressing as standard wound care. Results Wound healing outcome measures were recorded every week (at seven days) and compared at weeks 3, 6, and 9. There were no significant changes in the length and width of PUs between the groups till week 3. Significantly reduced length and width of NPD-treated PUs were found at week 6 (p=0.04) that further reduced at week 9 (p=0.001) as compared to standard wound care. Similarly, significant reduction in the depth of PUs was found in the NPD-treated group at week 9 (p<0.05). Exudate levels were significantly (p=0.001) lower in the NPD-treated group as compared to the standard wound care group from week 3 (2.96±0.21 vs 2.62±0.49); this difference continued through week 9 (1.35±0.75 vs 0.14±0.35). Disappearance of slough and formation of healthy granulation tissue was significantly higher in the NPD-treated PUs after week 6 (p=0.001). Conclusion NPWT may be the future of wound healing. As an alternative to the existing electrically powered NPWT systems, our NPD is safe, easy to apply, and efficacious in treating the PUs.

Effect of vitamin D supplementation on pain and physical function in patients with knee osteoarthritis (OA): an OA Trial Bank protocol for a systematic review and individual patient data (IPD) meta-analysis.

INTRODUCTION: Observational data suggest that vitamin D deficiency is associated with the onset and progression of knee osteoarthritis (OA). However, randomised controlled trials (RCTs) to date investigating the efficacy of vitamin D supplementation in knee OA have reported conflicting results. Further research is needed to clarify the effects of vitamin D on patient-reported outcomes and determine whether there are patient subgroups who may benefit from the supplementation. The aim of this individual patient data (IPD) meta-analysis is to identify patient-level predictors of treatment response to vitamin D supplementation on pain and physical function. METHODS AND ANALYSIS: A systematic literature search will be conducted for RCTs of vitamin D supplementation on knee OA. Authors of original RCTs will be contacted to obtain the IPD. The primary outcomes will include long-term (≥12 months) pain and physical function. Secondary outcomes will include medium-term (≥6 months and <12 months) and short-term (<6 months) pain and physical function, as well as patient global assessment, quality of life and adverse events. Potential treatment effect modifiers to be examined in the subgroup analyses include age, gender, body mass index, baseline knee pain severity and physical function, baseline vitamin D level, radiographic stage, presence of bone marrow lesions on MRI, presence of clinical signs of local inflammation and concomitant depressive symptoms. Both one-step and two-step modelling methods will be used to determine the possible modifiable effect of each subgroup of interest. ETHICS AND DISSEMINATION: Research ethical or governance approval is exempt for this study as no new data are being collected. This study will be the first IPD meta-analysis to clarify the effect of vitamin D supplementation on clinical symptoms in different subgroups of patients with knee OA. The findings will be disseminated through peer-review publications and conference presentations. PROSPERO REGISTRATION NUMBER: CRD42018107740.

Identifying the role of ASPN and COMP genes in knee osteoarthritis development.

BACKGROUND: Osteoarthritis (OA) is a common cause of musculoskeletal disability among elders and is characterized by late-onset degeneration of articular cartilage. OA affects various joints, commonly hand, knee, and hip, with clinical features that are unique to each joint. This study was initiated to identify and evaluate the role of the ASPN and COMP genes in the development of knee OA. METHODS: A case-control study was carried out involving 500 cases with knee OA (diagnosed by the American College of Rheumatology) and an equal number of healthy controls. Blood was drawn for genomic DNA isolation. PCR-RFLP and TaqMan assay methods were used to identify the SNPs. mRNA and protein expression of genes were carried out in peripheral blood lymphocytes (PBLs) by RT-PCR and Western immunoblotting. The data obtained were analyzed for the statistical significance between control and case groups. RESULTS: The variant genotype of ASPN and COMP genes was found to be present at a relatively higher frequency in cases than controls. RT-PCR and immunochemical studies revealed increased mRNA and protein expression of such gene in PBLs isolated from cases of knee OA as compared to healthy control. CONCLUSION: The allelic alteration in ASPN and COMP genes in knee OA cases points to the role of these genes in the development of knee OA. Further, increased mRNA and protein expression of ASPN and COMP in peripheral blood samples of patients with the disease suggest that expression profile of candidate gene could be used as a biomarker for predicting the development and progression of knee OA.

Augmentation of iNOS expression in myeloid progenitor cells expedites neutrophil differentiation.

Neutrophils play important role in immunity and inflammation through diverse mechanisms. Reports from this lab and others have demonstrated involvement of NO in neutrophil adhesion, chemotaxis, bacterial killing, reactive oxygen species generation, neutrophil extracellular traps' formation, and apoptosis. Constitutive expression of iNOS in human neutrophils has also been documented. The role of NO-iNOS in neutrophil differentiation however remains ill-defined. The present study was undertaken to understand the role of NO generated from iNOS in the neutrophil differentiation by using iNOS-overexpressing K562 cells (K562iNOS ) and iNOS-deficient murine progenitor cells (lineage negative cells; lin-ve ). We observed that iNOS overexpression led to increased neutrophilic differentiation in K562 cells; more specifically an early and accelerated neutrophilic differentiation was spotted in K562iNOS . These observations were further validated using iNOS knockout lin-ve cells or hematopoietic progenitor cells that exhibited delayed neutrophil differentiation in comparison to its wild-type counterpart. In addition, a significant increase in the gene expression of iNOS during neutrophilic differentiation of CD34+ hematopoietic stem and progenitor cells derived from human bone marrow further substantiates importance of iNOS in neutrophil differentiation. Moreover, a significant increase in NO generation during neutrophil differentiation was observed and enhanced neutrophil differentiation with NO donor was also observed, implying the importance of NO in neutrophil differentiation. Collectively, using alternative approaches, we demonstrated that neutrophil differentiation is significantly influenced by iNOS or NO, suggesting the possibility of exploiting this novel link for therapeutic aspects of NO generated from iNOS and neutrophil differentiation in hematopoiesis-related disorders.

Significance of methylation status of MASPIN gene and its protein expression in prognosis of gallbladder cancer.

AIM: We investigated prognostic significance of methylation status of MASPIN gene and its protein expression in normal subjects, cholelithiasis and gallbladder cancer (GBC) patients. METHODS: MASPIN protein expression and its promoter methylation in gallbladder tissue of cholelithiasis (n = 36), GBC (n = 46) and controls (n = 25) were investigated. Clinicopathological parameters and prognosis of patients with GBC were correlated with protein expression of MASPIN. Survival analysis of GBC patients was performed using Kaplan-Meier method. RESULTS: Significant increased (P < 0.0001) protein expression of MASPIN was observed in GBC as compared to cholelithiasis, whereas negligible expression was found in normal tissues. Methylation-specific PCR revealed statistical significant (P = 0.005) difference in methylation status of MASPIN promoter between gallstone and GBC patients. Significant association was observed between methylation profile with protein expression of MASPIN (P = 0.004), stage (P = 0.011) and cellular differentiation (P = 0.012) for GBC. Also, significant (P = 0.004) difference in survival was observed for malignant patients having demethylated MASPIN promoter. Further significant negative correlation (Pearson's coefficient [r] = -0.617; P < 0.0001) was observed between MASPIN expression and survival of cancer patients after surgery. Overall survival was significantly shorter (P ≤ 0.0001; hazard ratio [HR] for death = 2.84; 95% confidence interval [CI], 0.09865-0.3683) in patients of GBC with MASPIN expression ≥169.56 pg/mg (median survival; 10 months) with compared to those with expression <169.56 pg/mg (median survival; 16 months). CONCLUSION: Overexpression of MASPIN protein may play an important role in malignant progression and is correlated with a poor prognosis. Also MASPIN DNA methylation can be used as a novel therapeutic target for treating GBC.

Effectiveness of bone marrow-derived mononuclear stem cells for neurological recovery in participants with spinal cord injury: A randomized controlled trial.

BACKGROUND: Complete lesion after spinal cord injury (SCI) remains irreversible with little hope of neurological recovery. Newer interventions such as re-stimulation of damaged neurons using artificial agents and the use of stem cells for neuronal regeneration have shown promising results. AIM: This study was undertaken for evaluating the neurological status of acute SCI participants after stem cell augmentation and comparing them with other treatment methods. SETTING AND DESIGN: Randomized controlled trial in the northern Indian population. MATERIALS AND METHODS: A total 193 SCI participants of complete paraplegia with unstable T4-L2 injury having thoracolumbar injury severity score ≥4 were enrolled in this study. Participants were randomly allocated for three different treatment modalities, namely, conventional with stem cell augmentation (Group-1), conventional (Group-2), and conservative (Group-3). Neurological recovery after 1 year was evaluated through the ASIA Impairment Scale (AIS)-grading, sensory, and motor scores. STATISTICAL ANALYSIS: T-test for sensory-motor score analysis of each group and analysis of variance for comparison of same variables between the groups. RESULTS: After 1-year significant difference was observed in the AIS-grade, sensory and motor scores in-Group 1 (P < 0.001). In Group-1 versus 2, the mean difference at 1 year for AIS grade, sensory and motor scores were 0.40 (P = 0.010, 95% confidence interval [CI] 0.075-0.727), 8.52 (P = 0.030, 95% CI 0.619-16.419), and 4.55(P = 0.003, 95% CI 1.282-7.815), respectively. In Group-1 versus 3, 1.03, 19.02 and 7.22 (P < 0.001 for each of the parameters) and in Group-2 versus 3, 0.63 (P < 0.001), 10.49 (P = 0.009), and 2.68 (P = 0.019), respectively. CONCLUSIONS: Significant motor neurological recovery and AIS-grade promotion was observed in Group-1 as compared to Group-2 and 3.

Proton NMR based serum metabolic profile correlates with the neurological recovery in treated acute spinal cord injury (ASCI) subjects: A pilot study.

BACKGROUND: Acute Spinal Cord Injury (ASCI) is still having substantial morbidity and mortality despite of advanced therapeutics. Major obstacles are paucity of monitoring tools or biomarkers for severity determination, recovery and prognostication. A prospective case control pilot study with serum 1H NMR spectroscopic metabolic profiling was carried out to evaluate metabolites perturbations and its relationship with recovery and to see role of stem cells in facilitating neurological recovery. METHODOLOGY: Twenty subjects with ASCI were classified on the basis of therapeutic modality into surgical fixation alone (Group-1, n = 10), stem cell adjuvant (Group-2, n = 10) and healthy controls (Group-0, n = 10). Serum samples were collected at admission (baseline) and after six months (follow-up). NMR data of serum sample were quantified and subjected to Wilcoxon and ANOVA tests. Further validation was performed using supervised OSC-PCA and OPLS-DA by incorporating substantial control samples. RESULT: Twenty-eight metabolites were identified; well resolved resonances of fifteen metabolites were quantified wherein seven were statistically significant. Predominantly amino acids and ketone bodies played vital role in the differentiation of groups. CONCLUSIONS: Serum NMR spectroscopy reveals certain metabolites perturbations having clear correlation with pattern of recovery in treated ASCI subject. Stem cells treatment group had comparatively effective recovery.

Expression of Genes and Their Polymorphism Influences the Risk of Knee Osteoarthritis.

INTRODUCTION: Genetic factors including the level of expression of the fingerprint of genes involved in the development of bones and cartilage such as GDF-5 or ESR-α or CALM-1 are known to be strong determinants of the osteoarthritis (OA) in Caucasian and Oriental populations. Because of high prevalence of OA in Indian population and availability of limited genetic data, we determined whether similar genetic factors are involved in Indians as well. METHODS: A case control study was carried out involving 500 patients of knee OA and equal number of healthy controls. Genotyping analyses in whole blood, mRNA, and protein expressions in peripheral blood lymphocytes (PBLs) were performed using established protocols. RESULTS: Our results showed a significantly decreased level of mRNA and protein expressions for GDF-5, ESR-α, and CALM-1 genes in PBLs of OA cases when compared to healthy controls. The frequency of variant genotypes of these genes was also increased significantly in cases of OA compared to controls. CONCLUSION: Our results demonstrated that the decrease in expression of GDF-5, ESR-α, and CALM-1 in PBLs and association of polymorphism in these genes may be important in predicting the severity and thereby the progression of OA in Indian population.

Association between Single Nucleotide Polymorphisms of SMAD3 and BMP5 with the Risk of Knee Osteoarthritis.

INTRODUCTION: The role of genetic factors influencing osteoarthritis (OA) susceptibility is well documented and several candidate genes have been identified to be associated with it. Among these genes are Bone Morphogenetic Protein 5 (BMP5) and Smad family member 3 (SMAD3), all involved in Transforming Growth Factor (TGF) signaling pathway. The knee is the commonly affected joint, and knee OA has an especially high prevalence in Asian population. AIM: To investigate associations between Single Nucleotide Polymorphisms (SNPs) rs12901499 in SMAD3 and rs921126 in the BMP5 gene with knee OA susceptibility in and around Lucknow, Uttar Pradesh, India. MATERIALS AND METHODS: SNPs rs12901499 in SMAD3 and rs921126 in BMP5 were genotyped in patients with knee OA and age- sex matched OA-free controls from our population. A total of 450 patients with knee OA and 458 controls were enrolled in the study. Venous blood samples were obtained from all cases as well as controls for PCR-RFLP (Polymerase Chain Reaction- Restriction Fragment Length Polymorphism). Data was collected and entered in excel sheets. Statistical analyses of the data were performed using statistical software package SPSS version 16.0. Chi-square, Student's t-test and logistic regression tests were used to analyse the data. RESULTS: GA and GG genotypes of both SNPs (rs12901499 and rs921126), and variant G, were associated with a significantly increased risk of knee OA. A significantly increased risk of knee OA was associated with the genotype GG and GA of rs12901499 (p < 0.03 and p <0.004 respectively) and rs921126 (p< 0.0001 and p<0.001 respectively) compared with the AA genotype. In addition, those bearing at least one G allele (GG + GA) had a significantly increased risk of knee OA compared with those without the G allele (AA) in rs921126 (p< 0.0001). However, in rs12901499, significant association with the risk of knee OA was not found (p<0.4). On age and gender based stratification, the association between the risk of OA and rs921126 GG mutant compared with AA homozygotes was strong in both gender (adjusted OR= 2.93 for male and 2.25 for female) and in those aged >55 years (adjusted OR= 3.4), similarly in rs12901499, GG mutant compared with AA homozygote was strong in female (adjusted OR= 1.5) and in those aged >55 years (adjusted OR= 1.5). CONCLUSION: The results showed that both in SMAD3 rs12901499 and BMP5 921126, G allele is significantly associated with knee OA. A to G change and variant G genotype may contribute to knee OA risk in our study population of Lucknow.

Expression of MMP-8 in Pressure Injuries in Spinal Cord Injury Patients Managed by Negative Pressure Wound Therapy or Conventional Wound Care: A Randomized Controlled Trial.

PURPOSE: The purpose of this study was to assess the level of matrix metalloproteinase-8 (MMP-8) and wound-healing outcome measures (length, width, and depth, exudate amount, and tissue type) in pressure injuries (PIs) of spinal cord-injured patients treated with negative pressure wound therapy (NPWT) using a novel negative pressure device versus PI treated with wet to moist gauze (conventional wound care). DESIGN: Randomized controlled trial. SUBJECTS AND SETTING: Forty-four spinal cord-injured patients with stage 3 and 4 sacral PI participated in the study. The study setting was the Department of Orthopedic Surgery at King George's Medical University, in Lucknow, India. METHODS: Twenty two subjects were randomly allocated to undergo NPWT via a novel negative pressure device, and 22 participants received conventional wound dressing (wet to moist gauze dressings). Pressure injuries were treated for 9 weeks or until wound closure. Levels of MMP-8 were analyzed in the tissues of PIs at weeks 0, 3, 6, and 9 by enzyme-linked immunosorbent assay. RESULTS: Significantly lower levels of MMP-8 were observed in the NPWT group at week 6 and week 9. There were no significant changes in the length and width of PIs between the groups till week 3. Significant reduced length and width were observed in PIs of patients in the NPWT group at week 6 (P = .04) and week 9 (P = .001). Similarly, significant reduction in the depth of PIs was observed in the NPWT group at week 9 (P < .05). At the end of 9 week, levels of MMP-8 showed a positive correlation with reduction in the length, width, and depth of PIs in the NPWT group while in the conventional dressing group, negative correlation was observed in association with MMP-8 and the length, width, and depth of PIs. Exudate levels were significantly lower in the NPWT group compared with the conventional dressing group from week 3 (2.96 ± 0.21 vs 2.62 ± 0.49); this difference persisted through week 9 (1.35 ± 0.75 vs 0.14 ± 0.35). Conversion of slough into red granulation tissue was significantly higher in the NPWT group after week 6 (P = .001). CONCLUSION: Reduced levels of MMP-8 and an increased rate of healing were found in patients allocated to treatment with a novel negative pressure device as compared to wet to moist gauze conventional dressing. The novel NPWT device used in this study reduced exudate production and enhanced the rate of formation of red granulation tissue.

Proton NMR metabolic profiling of CSF reveals distinct differentiation of meningitis from negative controls.

BACKGROUND: Cerebrospinal fluid (CSF) is an essential bio-fluid of the central nervous system (CNS), playing a vital role in the protection of CNS and performing neuronal function regulation. The chemical composition of CSF varies during onset of meningitis, neurodegenerative disorders (positive controls) and in traumatic cases (negative controls). METHODS: The study design was broadly categorized into meningitis cases, negative controls and positive controls. Further differentiation among the three groups was carried out using Principal Component Analysis (PCA) followed by supervised Partial Least Square Discriminant Analysis (PLS-DA). RESULTS: The statistical analysis of meningitis vs. negative controls using PLS-DA model resulted in R2 of 0.97 and Q2 of 0.85. There was elevation in the levels of ketone bodies, total free amino acids, glutamine, creatine, citrate and choline containing compounds (choline and GPC) in meningitis cases. Similarly, meningitis vs. positive controls resulted in R2 of 0.80 and Q2 of 0.60 and showed elevation in the levels of total free amino acids, glutamine, creatine/creatinine and citrate in the meningitis group. Four cases of HIV were identified by PLS-DA model as well as by clinical investigations. CONCLUSION: On the basis of metabolic profile it was found that negative control CSF samples are more appropriate for differentiation of meningitis than positive control CSF samples.

CYR61-An angiogenic biomarker to early predict the impaired healing in diaphyseal tibial fractures.

BACKGROUND: Angiogenesis is a prerequisite for fracture repair, whereas insufficient blood supply is likely to result in impaired healing. In the present study, we aimed to determine the correlation of simple tibial fracture healing outcome with serial estimation of CYR61 expressions in the early phase of healing. METHODS: In total, 107 adult fractured patients and 97 healthy controls were analysed. Peripheral blood samples were taken from controls (at once) and fractured patients at 4th, 7th, 10th, 15th, 20th and 28th days of post-fracture follow-ups to quantify the CYR61 mRNA and protein expression by qRT-PCR and Western blotting assay, respectively. Clinic-radiological follow-up was done at 6th, 10th, 16th, 20th, and 24th weeks of post-fracture follow-ups using RUST scores to analyse the fracture healing progression and their final outcomes. RESULTS: By considering controls as Group I (n = 97), as per the clinico-radiological status at 24th week, fracture patients were divided into two groups: Group II (normal healing, n = 91) and Group III (impaired healing, n = 16). Both CYR61 mRNA and protein expressions were lower (baseline) in Group I than in Groups II and III; however, a significant difference was observed only with the Group II. In both groups, expressions of CYR61 mRNA as well as protein gradually upregulated from the baseline to a peak and then declined. Both, the CYR61 mRNA as well as protein expressions were significantly higher at all follow-ups in Group II than in Group III. Mean RUST scores between Group II and Group III showed a significant statistical difference at each follow-up. Significant correlation was found between the CYR61 expressions and the RUST score (fracture healing progression). CONCLUSION: We conclude that CYR61 expression provides an early prediction of the healing outcomes of simple diaphyseal tibial fractures. THE TRANSLATIONAL POTENTIAL OF THIS ARTICLE: Such an approach would benefit not only the patients' wellbeing but also the entire health care system in terms of the cost implications associated with long lasting treatment interventions and hospitalisation. However, the authors recommend further multicentric study with a large sample size to increase the validity, reliability, and generalisability of our observation and inferences.

A non-randomised, controlled clinical trial of an innovative device for negative pressure wound therapy of pressure ulcers in traumatic paraplegia patients.

The conventional methods of treatment of pressure ulcers (PUs) by serial debridement and daily dressings require prolonged hospitalisation, associated with considerable morbidity. There is, however, recent evidence to suggest that negative pressure wound therapy (NPWT) accelerates healing. The commercial devices for NPWT are costly, cumbersome, and electricity dependent. We compared PU wound healing in traumatic paraplegia patients by conventional dressing and by an innovative negative pressure device (NPD). In this prospective, non-randomised trial, 48 traumatic paraplegia patients with PUs of stages 3 and 4 were recruited. Patients were divided into two groups: group A (n = 24) received NPWT with our NPD, and group B (n = 24) received conventional methods of dressing. All patients were followed up for 9 weeks. At week 9, all patients on NPD showed a statistically significant improvement in PU healing in terms of slough clearance, granulation tissue formation, wound discharge and culture. A significant reduction in wound size and ulcer depth was observed in NPD as compared with conventional methods at all follow-up time points (P = 0·0001). NPWT by the innovative device heals PUs at a significantly higher rate than conventional treatment. The device is safe, easy to apply and cost-effective.

Does Serum Albumin Level Affect the Healing Outcomes of Simple Diaphyseal Tibial Fractures?

AIMS: To quantify the serum albumin level and its correlation with fracture healing progression and outcomes in adult patients. SETTINGS AND DESIGN: A prospective cohort study at an institutional trauma center. MATERIALS AND METHODS: A total of 50 adult patients with simple, fresh traumatic diaphyseal fractures of both bones of the leg managed conservatively were included in the study. Serum albumin was measured initially and at the 6th week postfracture. The clinico-radiological follow-up was done to analyze the fracture healing progression and their final outcomes, which were correlated with the quantified serum albumin level of the patients. STATISTICAL ANALYSIS USED: Student t-test, Mann-Whitney U-test and Pearson correlation coefficient. RESULTS: As per the last clinico-radiological follow-up at the 24th week, patients were grouped into two groups: Group I (normal union n = 38) and Group II (impaired healing n = 12). The mean serum albumin levels were significantly higher in Group I when compared to Group II. The association between the serum albumin level at baseline and at the 6th week was moderate. The best cut-off measure of serum albumin level was 3.45 g/dL, both at baseline and at the 6th week after fracture to predict the healing outcome. The correlation of serum albumin levels with fracture healing outcomes was statistically significant. CONCLUSION: Serum albumin is moderately associated with the baseline and 6th-week values and showed a positive correlation with the bony healing progression and may provide an early predictor of the healing outcomes of simple diaphyseal tibial fractures.