Conditioned medium-enriched umbilical cord mesenchymal stem cells: a potential therapeutic strategy for spinal cord injury, unveiling transcriptomic and secretomic insights.

INTRODUCTION: Spinal cord injury (SCI) leads to significant destruction of nerve tissue, causing the degeneration of axons and the formation of cystic cavities. This study aimed to examine the characteristics of human umbilical cord-derived mesenchymal stem cells (HUCMSCs) cultured in a serum-free conditioned medium (CM) and assess their effectiveness in a well-established hemitransection SCI model. MATERIALS AND METHODS: In this study, HUCMSCs cultured medium was collected and characterized by measuring IL-10 and identifying proteomics using mass spectroscopy. This collected serum-free CM was further used in the experiments to culture and characterize the HUMSCs. Later, neuronal cells derived from CM-enriched HUCMSC were tested sequentially using an injectable caffeic acid-bioconjugated gelatin (CBG), which was further transplanted in a hemitransection SCI model. In vitro, characterization of CM-enriched HUCMSCs and differentiated neuronal cells was performed using flow cytometry, immunofluorescence, electron microscopy, and post-transplant analysis using immunohistology analysis, qPCR, in vivo bioluminescence imaging, and behavioral analysis using an infrared actimeter. RESULTS: The cells that were cultured in the conditioned media produced a pro-inflammatory cytokine called IL-10. Upon examining the secretome of the conditioned media, the Kruppel-like family of KRAB and zinc-finger proteins (C2H2 and C4) were found to be activated. Transcriptome analysis also revealed an increased expression of ELK-1, HOXD8, OTX2, YY1, STAT1, ETV7, and PATZ1 in the conditioned media. Furthermore, the expression of Human Stem-101 confirmed proliferation during the first 3 weeks after transplantation, along with the migration of CBG-UCNSC cells within the transplanted area. The gene analysis showed increased expression of Nestin, NeuN, Calb-2, Msi1, and Msi2. The group that received CBG-UCNSC therapy showed a smooth recovery by the end of week 2, with most rats regaining their walking abilities similar to those before the spinal cord injury by week 5. CONCLUSIONS: In conclusion, the CBG-UCNSC method effectively preserved the integrity of the transplanted neuronal-like cells and improved locomotor function. Thus, CM-enriched cells can potentially reduce biosafety risks associated with animal content, making them a promising option for clinical applications in treating spinal cord injuries.

Effect of non-surgical periodontal therapy on CD14 + CD16+ monocyte counts in peripheral blood samples: a clinical interventional study.

Monocytes and their macrophage progeny are thought to be involved in tissue and alveolar bone destruction in periodontal disease. It has been documented that the proportion of (CD14 + CD16+) non-classical monocytes in the blood are elevated in chronic periodontitis;A total of 20 chronic generalized periodontitis patients who were otherwise healthy, were recruited for this study. At baseline and 3 weeks after non-surgical periodontal treatment, peripheral blood was obtained to assess the levels of C-reactive protein (CRP) and the proportion of monocyte subsets. Monocyte subsets were assessed using flow cytometry;The mean percentage of CD14 + CD16+ non-classical monocytes in the peripheral blood sample at baseline was 13.95 + 2.09, that reduced to 8.94 + 1.23 3 weeks after non-surgical treatment. A distinct significant reduction in the percentage of non-classical monocytes and a concomitant increase in classical monocytes were observed following periodontal treatment compared to baseline. There was a significant reduction in the all the periodontal parameters and CRP levels 3 weeks post non-surgical periodontal treatment. A positive correlation between CRP and percentage of non-classical monocytes was also observed; Periodontal treatment potentially modulates the host response effectively.

"Cancer Education on Wheels," a Novel Cost-Effective Method in Creating Awareness in the Community: a Pilot Study.

Cancer education raises disease awareness, the value of early identification and importantly the need for prompt screening and treatment when diagnosed. In this study, an effort was made to understand how well a unique cancer education program, "Cancer Education on Wheels," transfers knowledge in general public in society. The community was shown prerecorded cancer awareness videos using a TV monitor, CD player, and speaker system mounted on an eight-seat Toyota Innova. Before and after seeing the video presentation, consenting volunteers filled out questionnaires asking about demographics and understanding of cancer. Frequency and percentage calculations were done on the demographic information and a Wilcoxon signed-rank test was run on the overall subject score. Data was stratified based on demographic information and compared by applying Kruskal-Wallis test and Mann-Whitney test. p values under 0.05 were regarded as significant. A total of 584 people completed the pre-test and post-test questionnaires. The Wilcoxon signed-rank test revealed a difference between the pre-test and post-test (3.29 ± 2.48 vs. 6.78 ± 3.52; P = 0.0001). The pre-test results showed that volunteers between the ages of 18 and 30; men; students; urban residents; single volunteers; graduates; people who had known a person/or family member with cancer; and people who were aware of the suffering cancer causes had a high baseline knowledge of cancer (p = 0.015 to 0.001). The post-test results show that participants who had less baseline scores like housewives and unemployed people performed better (p = 0.006 to 0.0001). The findings unambiguously demonstrated that "Cancer Education on Wheels" was successful in raising participants' awareness of cancer signs and screening. Additionally, the findings showed that volunteers who were aged, married, housewives, and unemployed had higher scores. Most importantly, this approach to cancer education is simple to organize and carry out in a local setting. It is also affordable and simple to execute using readily available technological equipment and manageable logistics. According to the authors' knowledge, this is the first study to use "Cancer Education on Wheels" to spread awareness of cancer throughout the neighborhood especially in budget constrained areas.

Human gingival derived neuronal cells in the optimized caffeic acid hydrogel for hemitransection spinal cord injury model.

Spinal cord injury induces scar formation causes axonal damage that leads to the degeneration of axonal function. Still, there is no robust conceptual design to regenerate the damaged axon after spinal injury. Therefore, the present study demonstrates that human gingival derived neuronal stem cells (GNSCs) transplants in the injectable caffeic acid bioconjugated hydrogel (CBGH) helps to bridge the cavity and promote the engraftment and repopulation of transplants in the injured spinal tissue. Our study reports that the bioluminescence imaging in vivo imaging system (IVIS) provides a satisfactory progression in CBGH-GNSCs transplants compare to lesion control and CBGH alone. Immune regulators interleukin-6 (IL-6), tumor necrosis factor-α, neutrophil elastase are decreased, IL-10 is increased. Likewise, immunostaining (TAU/TUJ-1, SOX-2/NeuN, MAP-2/PSD93, NSE, S100b, and GFAP) shown repopulated cells. Also, TRA-1-81 expression confirms the absence of immune rejection in the CBGH-GNSCs transplants. However, locomotor recovery test, gene (IL-6, CASPASE3, p14-ARF, VEGF, LCAM, BDNF, NT3, NGN2, TrKc, FGF2, Sox-2, TUJ-1, MAP-2, Nestin, and NeuN) and protein expression (TAU, TUJ-1, SOX-2 MAP-2, PSD93, NeuN, TRA-1-81, GFAP, TAU, and MBP) shows functional improvements in the CBGH-GNSCs group. Further, GABA and glutamine level demonstrates the new synaptic vesicle formation. Hence, the CBGH scaffold enhances GNSCs transplants to restore the injured spinal tissue.

Salivary, Plasma, and Gingival Levels of Melatonin and TNF-α in Nonsmokers and Current Smokers with and without Periodontal Disease.

AIM: The aim of this study was to quantify the levels of gingival, salivary, and plasma melatonin and tumor necrosis factor-α (TNF-α) in healthy individuals and chronic generalized periodontitis patients with and without cigarette smoking habit and to investigate whether a relationship exists between melatonin and TNF-α levels in the samples. MATERIALS AND METHODS: Blood of 5 mL, 5 mL of saliva, and gingival tissue samples were obtained from 30 periodontally healthy individuals without smoking habit (HP), 30 nonsmoking patients with chronic generalized periodontitis (CP), 30 periodontally healthy individuals with current smoking habit (SHP), and 30 current smoker patients with chronic generalized periodontitis (SCP). The levels of melatonin and TNF-α in the samples were measured using commercially available enzyme-linked immunosorbent assay (ELISA) kit. The results obtained were statistically analyzed using SPSS statistical software (23.0 version). RESULTS: This study demonstrated the presence of melatonin and TNF-α in all the saliva, plasma, and gingival tissue samples. Gingival tissue melatonin levels were highest in the HP group and least in the SCP groups, while TNF-α levels were least in the HP group and highest in the SCP groups. No significant difference was observed between the groups with regard to salivary and plasma melatonin. An overall significant difference was also observed between the groups with regard to salivary TNF-α but not with regard to plasma TNF-α. Binary logistic regression analysis was carried out after dividing the study groups into current smokers and nonsmokers. Results revealed that a reduction in gingival melatonin and an increase in gingival TNF-α were associated with a transition from periodontal health to chronic generalized periodontitis in current smokers but not in nonsmokers. CONCLUSION: This study sheds light on the anti-inflammatory actions of melatonin in the gingival tissues in states of periodontal health and disease in current smokers. CLINICAL SIGNIFICANCE: Melatonin could be used as a supplement to boost anti-inflammatory mechanisms in periodontal therapy especially in cigarette smokers.

Human umbilical cord tissue stem cells and neuronal lineages in an injectable caffeic acid-bioconjugated gelatin hydrogel for transplantation.

Present-day scaffolds are useful in cell therapy to a reasonable extent, but in pursuit of improvising the scaffold to improve the outcome, we tested a new injectable caffeic acid-bioconjugated gelatin hydrogel scaffold (CBGH; with tunable stiffness -10%). Two-dimensional (2D) form of human umbilical cord tissue-derived mesenchymal stem cells (HUCMSCs) culture performed based on our previously reported methods and characterized by using multipotent and pluripotent analysis. In addition, neurogenesis was induced in the presence of retinoic acid or neural growth factor or epidermal growth factor categorized by neuronal markers. The viability, proliferation rate, and vascular endothelial growth factor expression of HUCMSCs increased significantly in the CBGH scaffold. In addition, there was an increase in CD90 and TRA-1-81 phenotypic expressions and SOX-2, MAP-2, TAU, NeuN, and NF, which confirmed the neurogenesis of encapsulated HUCMSCs. Topographical elucidation by scanning electron microscopy data showed that the HUCMSCs proliferated and migrated inside the construct. Hematoxylin and eosin staining demonstrated a more viable structural pattern and cresyl violet staining showed the Nissl synthesis, confirming the presence of functional neurons in the encapsulated form. The molecular-level analysis further substantiated that HUCMSCs cultured in CBGH expressed significantly greater upregulation of stemness, neuronal genes, and protein expression compared with the adherent culture. Correspondingly, this is the first time that we have measured the fluorescence intensity variation of the HUCMSCs-stained cell segmentation process using customized MATLAB code execution to reduce the background noise and autofluorescence. We conclude that this novel CBGH scaffold increases the viability, proliferation, stemness, and also neuronal transdifferentiation of HUCMSCs in a three-dimensional culture than the 2D plastic adherent culture.

Passage-dependent expression of STRO-1 in human gingival mesenchymal stem cells.

The expression of STRO-1, the essential mesenchymal stem cell marker, was found to decrease with advancing passages in few tissues. Because STRO-1 was identified and isolated from human gingiva, we were interested to know its status after a few passages. Human gingival mesenchymal stem cells (HGMSCs) were isolated from human gingiva. Flow cytometry was carried out with STRO-1, mesenchymal stem cell (MSC) positive marker CD73, and negative marker CD34/CD45. Samples were also subjected to CD90 and STRO-1 immunofluorescence staining. Gene expression was carried out for transcription factors OCT-4, NANOG, and NESTIN. The results showed a gradual decrease in STRO-1 and transcription factor expression with an increase in passage numbers. MSC positive marker CD73 was consistently expressed in all the passages. Negative markers were absent in all the passages. We conclude that STRO-1 may be a useful marker to isolate undifferentiated (potent) mesenchymal cells from gingiva.

The Relationship Between Self-Esteem, Emotional Intelligence, and Empathy Among Students From Six Health Professional Programs.

Theory: Psychosocial skills such as communication, empathy, and emotional intelligence are now considered key attributes of health professionals. Self-esteem is another important construct that is less well studied. Self-esteem is important because low levels have been linked to depression, suicide, and eating disorders. Given that health professional students experience high levels of stress and are at increased risk for similar psychopathology, self-esteem may be an important variable in student well-being and performance after graduation. Hypotheses: This study sought to explore self-esteem during students' 1st year of training hypothesizing that several would demonstrate low self-esteem. It is also hypothesized that emotional intelligence and empathy would be associated with self-esteem. Method: A cross-sectional survey was conducted, and data were gathered from dental, medical, nursing, optometry, pharmacy and veterinary students. Self-report questionnaires assessing self-esteem, emotional intelligence, and empathy were completed and demographic information was collected. Scores were calculated and differences between groups analyzed with analysis of variance and chi-square testing. Pearson's correlation was used to assess associations between the constructs. Results: The mean self-esteem score was 26.2 ± 2.3 but 21% of the sample evidenced low self-esteem. There was no difference in the proportion of students demonstrating low self-esteem among programs. Gender did not have a significant effect on self-esteem scores, though ethnicity did. Emotional intelligence scores were higher among male individuals than among female. Emotional intelligence and empathy showed a small association with self-esteem. Conclusions: A significant proportion of health professional students suffer from low self-esteem during their 1st year of study. Such students may be more susceptible to the stresses associated with study and the development of psychopathology. More research needs to be conducted to explore the relationships between self-esteem, emotional intelligence, and empathy with a view to strengthening training in these areas and managing the challenges faced by health professional students.

Survey of multinational surgical management practices in tetralogy of Fallot.

BACKGROUND: A wide variety of surgical strategies are used in tetralogy of Fallot repair. We sought to describe the international contemporary practice patterns for surgical management of tetralogy of Fallot. METHODS: Surgeons from 18 international paediatric cardiac surgery centres (representing over 1800 tetralogy of Fallot cases/year) completed a Research Electronic Data Capture-based survey. Participating countries include: China (4), India (2), Nepal (1), Korea (1), Indonesia (1), Saudi Arabia (3), Japan (1), Turkey (1), Australia (1), United States of America (2), and Canada (1). Summary measures were reported as means and counts (percentages). Responses were weighted based on case volume/centre. RESULTS: Primary repair is the prevalent strategy (83%) with variation in age at elective repair (range). Approximately 47% of sites use patient age as a factor in determining the strategy, with age 90% of all trans-annular repairs. CONCLUSIONS: In this cohort representing 11 countries, there is variation in tetralogy of Fallot surgical management with no consensus on standard of practice. A large international prospective cohort study would allow analysis of impact of underlying anatomy and repair strategy on early and late outcomes.

Transdifferentiation of human gingival mesenchymal stem cells into functional keratinocytes by Acalypha indica in three-dimensional microenvironment.

Gingival tissue is reportedly a promising, easily accessible, abundant resource of mesenchymal stem cells (MSC) for use in various tissue engineering strategies. Human gingival MSC (HGMSCs) were successfully isolated from gingival tissue and characterized. To analyze in a two-dimensional form, HGMSCs were cultured with basal medium and induced with 25 µg/ml of Acalypha indica. Quantitative real-time polymerase chain reaction (qPCR) and western blot analysis showed the presence of keratinocyte-specific markers, including cytokeratin-5 and involucrin. To further assess its capability for stratification akin to human keratinocytes, HGMSCs were encapsulated in a HyStem® -HP Cell Culture Scaffold Kit and cultured in the presence of A. indica. Calcein AM staining indicated that the HyStem® -HP Scaffold Kit has excellent biocompatibility. Immunofluorescence and qPCR analysis revealed the presence of keratinocyte-specific markers. The study concluded that the three-dimensional microenvironment is a novel method for inducing epidermal differentiation of HGMSCs to engineer epidermal substitutes with the help of A. indica, which provides an alternative strategy for skin tissue engineering.

Gingival spheroids possess multilineage differentiation potential.

Recently studies have demonstrated HGMSCs as ideal candidates for regenerative study. Interestingly we found that HGMSCs derived spheroids are more potent and maintain the properties of stemness convincingly compared to conventional culture methods. During the culture, GMSCs instinctively accumulated into spheroids and display multipotent STRO-1 and Vimentin-positive cells. Reduced phenotypic expression of CD73, CD105, and elevated expression STRO-1 and CD-34. Pluripotent nature of S-GMSCs putatively shown the expression of OCT4A, NANOG, SOX-2, SSEA4, TRA-1-60, and TRA-181. Also, levels of protein are much higher in spheroid than dissociated culture. On endothelial induction, spheroid differentiated and developed a vascular structure with positive expression of CD31 and on neuronal induction showed positivity for TUJ1 and E-Cadherin. Importantly, undifferentiated state of S-GMSCs exhibited significant upregulation of aforementioned pluripotent genes and lack of pro-inflammatory cytokines IL-6 and amplified ARF signal confirming that the spheroids are not teratoma formation. However, higher of CAP1, CP, TGFß, OPN, PPARÉ£, TUJ1, and NESTIN expression observed in spheroids, and minimal expression of the same markers were observed in adherent GMSCs respectively. Ahead of dissociated gingival culture, spheroid provides enhanced viable, pluripotent, and multilineage ability. This study suggested that S-GMSCs increased the chances of therapeutic efficacy in the regenerative applications.

4PBA strongly attenuates endoplasmic reticulum stress, fibrosis, and mitochondrial apoptosis markers in cyclosporine treated human gingival fibroblasts.

Cyclosporine induces overgrowth of human gingiva. Previously we have shown (i) cyclosporine-inducing ER stress in human gingival fibroblasts (HGF), (ii) increased matrix protein expression, and (iii) interference with mitochondrial pro- and anti-apoptotic factors. This study was undertaken to assess the effects of melatonin (an antioxidant), 4PBA (an ER stress inhibitor), and simvastatin on the expression of ER Stress markers as well as on matrix and mitochondrial markers. HGF incubated with cyclosporine, or without melatonin/4PBA/statin. After 24 hr of incubation, mRNA expression of ER stress markers (GRP78, CHOP, XBP1, and XBPs) and matrix protein markers (like α-SMA, VEGF, TGF-ß, CTGF), and mitochondrial apoptosis markers estimated and compared with housekeeping gene GAPDH. Compared to the control cyclosporine significantly augmented ER Stress and matrix proteins, which decreased significantly with the use of melatonin, 4PBA, and simvastatin. The mitochondrial proapoptotic molecule cyclophilin D, as well as Bcl2 expression also decreased after PBA treatment, paralleling an increase in cytochrome c expression. The effect of 4PBA was much more pronounced than the influence of other two. In conclusion, 4PBA could be a viable therapeutic option for drug-induced gingival overgrowth.

Differential expression of microRNAs let-7a, miR-125b, miR-100, and miR-21 and interaction with NF-kB pathway genes in periodontitis pathogenesis.

Periodontitis is a chronic inflammatory disease which is caused by destruction of the tissues that surrounds and supports the tooth. Deregulation of microRNAs has been reported to cause several inflammatory diseases such as autoimmune disease, chronic periodontitis, and cancer. In the present study, we have investigated the expression pattern of microRNAs let-7a, miR-125b, miR-100, miR-21, and RNA-binding protein LIN-28A among healthy individuals and chronic periodontitis patients. Total RNA was isolated from gingival tissue samples collected from 100 healthy individuals and 100 chronic periodontitis patients. The expression of microRNAs and LIN-28 was performed by qPCR. Target prediction for the microRNAs was done using miRWalk and miRTarbase online databases and the experimentally validated targets were analyzed for their molecular function, biological processes, and related pathways using gProfiler software. The expression analysis revealed that let-7a and miR-21 were upregulated, whereas, miR-100, miR-125b, and LIN-28 were down regulated. The age dependent expression analysis revealed that the expression levels of all the microRNAs and LIN-28 were found to increase with age (more than 50 years), thereby suggesting an increased risk to chronic periodontitis. Among the various targets predicted using miRWalk and miRTarbase databases, NFKB was found to be a common target among all the four microRNAs. gProfiler revealed several functions such as NF-ĸB signaling pathway, cytokine-cytokine receptor interaction, osteoclast differentiation, etc., all of which specific to inflammation and periodontitis.

Osteogenic differentiation of human gingival mesenchymal stem cells by Aristolochia bracteolata supplementation through enhanced Runx2 expression.

The phenotypic characteristics of human gingival derived mesenchymal stem cells (HGMSCs) on induction with total methanol extract of Aristolochia bracteolata have been evaluated. HGMSCs were cultured in control and two different induction medium: Control medium (basal medium), OM1 (Standard induction medium), and OM2 (100 µg/ml of A. bracteolata). Osteogenic differentiation of the cultured cells was assessed by studying the calcium deposition and osteoblastic gene expression. OM2 medium showed an enhanced osteogenic differentiation potential than OM1 as measured by increased calcium deposition and elevated expression of Runx2, osteopontin, osteonectin, osteocalcin, Collagen type I, and ALP levels in comparison with OM1 differentiated cells. We conclude that at 100 µg/ml A. bracteolata has induced HGMSC differentiation into osteogenic lineage consequent to enhanced Runx2 expression and related osteogenic genes.

Comparision of Gingival and Umbilical Cord Stem Cells Based on Its Modulus and Neuronal Differentiation.

The availability of Human Umbilical Cord-derived Mesenchymal Stem Cells (HUCMSCs) from a single sex being a major limitation for the utilization of a potential stem cell, it is highly desirable to utilize, an autogenous pluripotent cell with desirable biological and mechanical properties in clinical situations. Comparison of Human Gingival Mesenchymal Stem Cells (HGMSCs) with HUCMSCs demonstrates; MSCs derived from gingiva have higher proliferation rate and higher population doubling time than Umbilical Cord. Unlike HUCMSCs, immunofluorescence studies showed the presence of pluripotency markers OCT-4 and NANOG predominantly in the cytoplasm of HGMSCs which was confirmed by Western blot. The mechanical property, such as modulus of elasticity of HGMSCs, is on par with HUCMSCs, but the surface roughness found to be lesser in HGMSCs, which may suggest HGMSCs greater adhesive property to the extracellular matrix. There is a marginal difference in the neuronal differentiation rate between HGMSCs and HUCMSCs; both the cells expressed positivity for several neuronal lineage markers. Hence, HGMSCs represent an autogenous source of mesenchymal stem cells, which are easy to procure with least morbidity, multipotent in nature with desirable biological, and mechanical properties, probably an ideal candidate for clinical applications. J. Cell. Biochem. 118: 2000-2008, 2017. © 2017 Wiley Periodicals, Inc.

Cyclosporine-A induces endoplasmic reticulum stress and influences pro-apoptotic factors in human gingival fibroblasts.

Cyclosporine-A (CsA) induces gingival overgrowth. Cyclosporine's anti-apoptotic activity in human gingival fibroblast is due to desensitization of mitochondrial permeability transition pore (MPTP) and augmentation of anti-apoptotic, Bcl2. Alternative mechanisms of apoptosis exist involving enzymes like calcium-dependent Calpain and signaling events related to apoptosis, like Glycogen synthase kinase 3ß (GSK3ß) and protein kinase A (PKA). Cyclosporine-A in renal tubular cells induces endoplasmic reticulum stress (ER stress) which has not been explored in gingival overgrowth. Hence, this study was carried out to assess the influence of Cyclosporine-on ER stress and on the alternate anti-apoptotic mechanisms. Human gingival fibroblasts were treated with CsA, and expression of ER stress markers, such as binding immunoglobulin protein and CCAAT/enhancer-binding protein homologous protein (CHOP), MPTP, and expression of Calpain & GSK3ß /PKA were estimated. The results showed CsA-added fibroblast significantly increasing the expression of Endoplasmic reticulum stress markers. Contrary to usual ER stress outcome of apoptosis, we observed Cyclosporine's anti-apoptotic action in spite of augmented ER stress markers. We conclude that CsA's independent action on different organelles may alter the conventional outcome of ER stress.

Glucose sensing in oral mucosa simulating phantom using differential absorption based frequency domain low-coherence interferometry.

The superluminescent diode based differential absorption frequency domain low-coherence interferometry (FD-DALCI) technique is proposed and demonstrated for sensing physiological concentrations of glucose (0-250 mg/dl) in oral mucosa simulating phantoms (intralipid of concentrations 0.25-0.50%) with wavelengths at 1589 and 1310 nm. The proposed technique allows simultaneous measurements of refractive index based spectral shift and estimation of physiological concentration of glucose in intralipid with scattering characteristics using the differential absorption approach. The sensitivity of the glucose concentration obtained by spectral shift measurement was ≈0.016 nm/(mg/dl), irrespective of the intralipid concentration. The resolution of the glucose level was estimated to be ≈15 mg/dl in 0.25% intralipid and ≈19 mg/dl in 0.5% intralipid using the FD-DALCI technique.

Rare associations of tetralogy of Fallot with anomalous left coronary artery from pulmonary artery and totally anomalous pulmonary venous connection.

We describe the cases of two patients with tetralogy of Fallot, aged 4 years and 8 months, who were incidentally detected to have concomitant anomalous left coronary artery from pulmonary artery and total anomalous pulmonary venous connection, respectively, on preoperative imaging. They underwent surgical correction with good mid-term outcomes. In this study, we discuss the embryological basis, physiological effects, and review the literature of these two unusual associations. Awareness of these rare associations will avoid missed diagnoses and consequent surgical surprises.

Elevated levels of cyclooxygenase 1 and 2 in human cyclosporine induced gingival overgrowth.

OBJECTIVE: This study was carried out to immuno-localize and estimate the levels of cyclooxygenase 1 and 2 in human gingival tissue samples from healthy individuals, chronic periodontitis patients and patients with cyclosporine induced gingival overgrowth. METHODS: Group I consisted of individuals with healthy gingiva (n=6), Group II - cyclosporine induced gingival overgrowth (n=9) and Group III - chronic periodontitis patients (n=6). Gingival tissue samples were collected from subjects of all the three groups. COX-1, COX-2 levels were estimated in tissue homogenates by enzyme activity assay. Immuno-localization for COX-1 and COX-2 was also done in sections of gingival tissue. RESULTS: The study results demonstrated a significantly higher mean levels of COX-1 and 2 in drug induced gingival overgrowth samples (p<0.05). COX-1 and COX-2 was localized to epithelium and connective tissue in human gingival tissue sections from cyclosporine induced gingival overgrowth. CONCLUSION: Cyclooxygenase enzymes appear to be potential mediators involved in the pathogenesis of cyclosporine induced gingival overgrowth.