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1.  Benign Pairs: A Significant Entity in Aspiration Cytology Smears From Breast Lesions 
Fine needle aspiration cytology (FNAC) is a reliable diagnostic tool used to diagnose breast lesions preoperatively. However, FNAC is also associated with diagnostic pitfalls. Further studies are needed to improve its diagnostic efficacy. We noticed ovoid, bare nuclei arranged in closely touching diads (benign pairs) in a significant number of cytology smears. This prompted us to assess their diagnostic utility.
This was a prospective study conducted in Sarojini NaiduMedical College, Agra, India. Quantitative estimation of benign pairs per 1000 ductal cells in at least 20 high power field was attempted in cytology smears of 128 cases.
The average number of pairs in benign and malignant lesions was calculated as 7.07 + 5.96 and 0.28 + 0.78, respectively. Statistical analysis showed a significant difference between the number of pairs in benign and malignant cases (P < 0.0005).
Quantitative estimation of benign pairs is helpful in distinguishing benign from malignant cases.
PMCID: PMC3855199  PMID: 24324356
benign pairs; breast cytology; fine needle aspiration cytology
2.  Shear Strength Behavior of Human Trabecular Bone 
Journal of biomechanics  2012;45(15):2513-2519.
The shear strength of human trabecular bone may influence overall bone strength under fall loading conditions and failure at bone-implant interfaces. Here, we sought to compare shear and compressive yield strengths of human trabecular bone and elucidate the underlying failure mechanisms. We analyzed 54 specimens (5-mm cubes), all aligned with the main trabecular orientation and spanning four anatomic sites, 44 different cadavers, and a wide range of bone volume fraction (0.06–0.38). Micro-CT-based non-linear finite element analysis was used to assess the compressive and shear strengths and the spatial distribution of yielded tissue; the tissue-level constitutive model allowed for kinematic non-linearity and yielding with strength asymmetry. We found that the computed values of both the shear and compressive strengths depended on bone volume fraction via power law relations having an exponent of 1.7 (R2=0.95 shear; R2=0.97 compression). The ratio of shear to compressive strengths (mean ± SD, 0.44 ± 0.16) did not depend on bone volume fraction (p=0.24) but did depend on microarchitecture, most notably the intra-trabecular standard deviation in trabecular spacing (R2=0.23, p<0.005). For shear, the main tissue-level failure mode was tensile yield of the obliquely oriented trabeculae. By contrast, for compression, specimens having low bone volume fraction failed primarily by large-deformation-related tensile yield of horizontal trabeculae and those having high bone volume failed primarily by compressive yield of vertical trabeculae. We conclude that human trabecular bone is generally much weaker in shear than compression at the apparent level, reflecting different failure mechanisms at the tissue level.
PMCID: PMC3462285  PMID: 22884967
shear; finite element; trabeculae; microarchitecture; yield
3.  Passive Smoking Impairs Histone Deacetylase-2 in Children With Severe Asthma 
Chest  2013;145(2):305-312.
Parental smoking is known to worsen asthma symptoms in children and to make them refractory to asthma treatment, but the molecular mechanism is unclear. Oxidative stress from tobacco smoke has been reported to impair histone deacetylase-2 (HDAC2) via phosphoinositide-3-kinase (PI3K)/Akt activation and, thus, to reduce corticosteroid sensitivity. The aim of this study was to investigate passive smoking-dependent molecular abnormalities in alveolar macrophages (AMs) by comparing passive smoke-exposed children and non-passive smoke-exposed children with uncontrolled severe asthma.
BAL fluid (BALF) was obtained from 19 children with uncontrolled severe asthma (10 non-passive smoking-exposed subjects and nine passive smoking-exposed subjects), and HDAC2 expression/activity, Akt/HDAC2 phosphorylation levels, and corticosteroid responsiveness in AMs were evaluated.
Parental smoking reduced HDAC2 protein expression by 54% and activity by 47%, with concomitant enhancement of phosphorylation of Akt1 and HDAC2. In addition, phosphorylation levels of Akt1 correlated positively with HDAC2 phosphorylation levels and negatively with HDAC2 activity. Furthermore, passive smoke exposure reduced the inhibitory effects of dexamethasone on tumor necrosis factor-α-induced CXCL8 release in AMs. There were relatively higher neutrophil counts and CXCL8 concentrations in BALF and lower Asthma Control Test scores compared with non-passive smoke-exposed children with uncontrolled severe asthma.
Passive smoking impairs HDAC2 function via PI3K signaling activation, which could contribute to corticosteroid-insensitive inflammation in children with severe asthma. This novel mechanism will be a treatment target in children with severe asthma and stresses the need for a smoke-free environment for asthmatic children.
PMCID: PMC3913299  PMID: 24030221
4.  The Diagnostic Role of the Marginal Vacuoles in FNAC of Solitary Thyroid Nodules 
Context: FNAC thyroid is more sensitive as well as specific in segregating neoplastic and non-neoplastic thyroid lesions. Identification of predominant cell pattern, cell morphology and background details in FNAC smears play an important role in categorisation of various thyroid lesions with accuracy. Marginal vacuoles (MVs) have been described as irregular cytoplasmic vacuoles with largely unstained central area. MVs, although suggestive of thyrotoxicosis, are non-specific, as they are also encountered in non- toxic goitre, Hashimoto’s thyroiditis and follicular carcinoma. This cytologic finding therefore deserves further study to precisely delineate its utility.
Aims & Objective: To investigate whether MVs have a diagnostic role in FNAC of solitary thyroid nodule
Setting & Design: This study was conducted on 40 patients who presented with solitary thyroid nodule.
Methods & Material: Forty patients having solitary thyroid nodule were subjected to FNAC on an outpatient basis. Their cytomorphological features were studied with special attention to presence of MVs. MVs if present were then graded as no/ scant (gradeI ), moderate (gradeII) and abundant (gradeIII).
Results and Conclusion: Majority (77.5%) of nodular goitres had insignificant MVs (gradeI) in their smears. Significant MVs (gradeII+III) were limited to autoimmune thyroiditis presenting as solitary nodule and follicular neoplasms. Abundant MVs/ gradeIII were seen in follicular neoplasm only. MVs were not present in rest of neoplastic goitres. This implies that MVs are a marker of follicular differentiation.
PMCID: PMC3592293  PMID: 23542577
Thyroid; Marginal Vacuoles; Solitary thyroid nodule
5.  Cytomorphologic significance of marginal vacuoles in diffuse thyroid enlargements 
Fine needle aspiration cytology (FNAC) of the thyroid is an established first-line test for thyroid lesions. Marginal vacuoles (MVs) have been associated with hyperactivity of the thyroid, but some studies have pointed towards their nonspecific status.
To assess the presence of MVs in diffuse thyroid enlargements and evaluate the strength of correlation between MVs, levels of thyroid hormone and cytological diagnosis.
Materials and Methods:
Ninety-seven cases of diffuse thyromegaly were studied. Cytomorphological features were examined with special attention to MVs. MVs were graded as scant, moderate and abundant. Hormonal status of the patients was recorded. The presence and grading of MVs was correlated with cytological diagnosis and hormonal status. The strength of association was studied by applying the Chi-square test and test of proportion; a P ≤ 0.05 was considered significant.
Abundant MVs were not associated with hypothyroidism in this study; 79% of these cases were hyperthyroid. The correlation between moderate/insignificant MVs and functional status of the thyroid gland was inconclusive. Further, abundant MVs in thyroid FNACs were seen in cases of primary hyperplasia and Hashimoto's thyroiditis. There was a significant correlation between the presence of abundant/moderate MVs and primary hyperplasia and their absence in colloid goiter (P = 0.01 and 0.004, respectively).
A significant association was found between abundant MVs and a hyperthyroid state. Moderate/absent MVs in diffuse goiters were not found to correlate with thyroid function. Thus, all diffuse goiters with prominent MVs require hormonal evaluation to rule out hyperfunction of the thyroid.
PMCID: PMC3701337  PMID: 23833403
Marginal vacuoles; thyroid fine needle aspiration cytology; thyroid hormones
6.  Relationship between Serum Vitamin D, Disease Severity, and Airway Remodeling in Children with Asthma 
Little is known about vitamin D status and its effect on asthma pathophysiology in children with severe, therapy-resistant asthma (STRA).
Relationships between serum vitamin D, lung function, and pathology were investigated in pediatric STRA.
Serum 25-hydroxyvitamin D [25(OH)D3] was measured in 86 children (mean age, 11.7 yr): 36 with STRA, 26 with moderate asthma (MA), and 24 without asthma (control subjects). Relationships between 25(OH)D3, the asthma control test (ACT), spirometry, corticosteroid use, and exacerbations were assessed. Twenty-two of 36 children with STRA underwent fiberoptic bronchoscopy, bronchoalveolar lavage, and endobronchial biopsy with assessment of airway inflammation and remodeling.
Measurements and Main Results
25(OH)D3 levels (median [IQR]) were significantly lower in STRA (28 [22–38] nmol/L) than in MA (42.5 [29–63] nmol/L) and control subjects (56.5 [45–67] nmol/L) (P < 0.001). There was a positive relationship between 25(OH)D3 levels and percent predicted FEV1 (r = 0.4, P < 0.001) and FVC (r = 0.3, P = 0.002) in all subjects. 25(OH)D3 levels were positively associated with ACT (r = 0.6, P < 0.001), and inversely associated with exacerbations (r=−0.6, P < 0.001) and inhaled steroid dose (r=−0.39, P = 0.001) in MA and and STRA. Airway smooth muscle (ASM) mass, but not epithelial shedding or reticular basement membrane thickness, was inversely related to 25(OH)D3 levels (r=−0.6, P = 0.008). There was a positive correlation between ASM mass and bronchodilator reversibility (r = 0.6, P = 0.009) and an inverse correlation between ASM mass and ACT (r = −0.7, P < 0.001).
Lower vitamin D levels in children with STRA were associated with increased ASM mass and worse asthma control and lung function. The link between vitamin D, airway structure, and function suggests vitamin D supplementation may be useful in pediatric STRA.
PMCID: PMC3471128  PMID: 21908411
vitamin D; asthma; remodeling; airway smooth muscle; pediatrics
7.  1α,25-Dihydroxyvitamin D3 promotes CD200 expression by human peripheral and airway-resident T cells 
Thorax  2012;67(7):574-581.
CD200, a cell-surface immunoglobulin-like molecule expressed by immune and stromal cells, dampens the pro-inflammatory activity of tissue-resident innate cells via its receptor, CD200R. This interaction appears critical for peripheral immune tolerance, particularly in the airways where excessive inflammation is undesirable. Vitamin D contributes to pulmonary health and promotes regulatory immune pathways, therefore its influence on CD200 and CD200R was investigated.
CD200 and CD200R expression were assessed by qPCR and immunoreactivity of human lymphoid, myeloid and epithelial cells following 1α,25-dihydroxyvitamin D3 (1α,25VitD3) exposure in vitro and in peripheral T cells following 1α,25VitD3 oral ingestion in vivo. The effect of 1α25VitD3 was also assessed in human airway-resident cells.
1α25VitD3 potently upregulated CD200 on peripheral human CD4+ T cells in vitro, and in vivo there was a trend towards upregulation in healthy, but not asthmatic individuals. CD200R expression was not modulated in any cells studied. CD200 induction was observed to a lesser extent in CD8+ T cells and not in B cells or airway epithelium. T cells isolated from the human airway also responded strongly to 1α25VitD3 to upregulate CD200.
The capacity of 1α,25-dihydroxyvitamin D3 to induce CD200 expression by peripheral and respiratory tract T cells identifies an additional pathway via which vitamin D can restrain inflammation in the airways to maintain respiratory health.
PMCID: PMC3471129  PMID: 22334534
8.  The role of 1α,25-dihydroxyvitamin D3 and cytokines in the promotion of distinct Foxp3+ and IL-10+ CD4+ T cells 
European journal of immunology  2012;42(10):2697-2708.
1α,25-Dihydroxyvitamin D3 (1α25VitD3) has potent immunomodulatory properties. We have previously demonstrated that 1α25VitD3 promotes human and murine IL-10-secreting CD4+ T cells. Because of the clinical relevance of this observation, we characterized these cells further and investigated their relationship with Foxp3+ regulatory T (Treg) cells. 1α25VitD3 increased the frequency of both Foxp3+ and IL-10+ CD4+T cells in vitro. However, Foxp3 was increased at high concentrations of 1α25VitD3 and IL-10 at more moderate levels, with little coexpression of these molecules. The Foxp3+ and IL-10+ T-cell populations showed comparable suppressive activity. We demonstrate that the enhancement of Foxp3 expression by 1α25VitD3 is impaired by IL-10. 1α25VitD3 enables the selective expansion of Foxp3+ Treg cells over their Foxp3− T-cell counterparts. Equally, 1α25VitD3 maintains Foxp3+ expression by sorted populations of human and murine Treg cells upon in vitro culture. A positive in vivo correlation between vitamin D status and CD4+Foxp3+ T cells in the airways was observed in a severe pediatric asthma cohort, supporting the in vitro observations. In summary, we provide evidence that 1α25VitD3 enhances the frequency of both IL-10+ and Foxp3+ Treg cells. In a translational setting, these data suggest that 1α25VitD3, over a broad concentration range, will be effective in enhancing the frequency of Treg cells.
PMCID: PMC3471131  PMID: 22903229
1α,25-Dihydroxyvitamin D3; Asthma; Immune regulation; Regulatory T cells
10.  Multiple Pulp Stones in Primary and Developing Permanent Dentition: A Report of 4 Cases 
Case Reports in Dentistry  2012;2012:408045.
Pulp stones are foci of calcification or discrete calcifications in the dental pulp. They are frequently found on bitewing and periapical radiographs, but their occurrence in entire dentition is unusual. We are reporting four cases in which the occurrence of pulp stones ranged from their presence in just primary teeth (Cases 1 and 2) to involvement of young permanent teeth also (Case 3) and even unerupted permanent teeth (Case 4). In all the cases, dental, medical, and family histories as well as the findings from the clinical examination of the patient were not contributory. Histopathological report revealed true denticle. Metabolic evaluation of patients through liver function test, kidney function test, and blood investigation did not show any metabolic disorders. Patients were also evaluated for any systemic, syndromic, or genetic involvement, but this was also noncontributing. Therefore, it is suggested that these unusual cases may be of idiopathic origin.
PMCID: PMC3434379  PMID: 22970392
11.  Relationship of Nitrogen Use Efficiency with the Activities of Enzymes Involved in Nitrogen Uptake and Assimilation of Finger Millet Genotypes Grown under Different Nitrogen Inputs 
The Scientific World Journal  2012;2012:625731.
Nitrogen responsiveness of three-finger millet genotypes (differing in their seed coat colour) PRM-1 (brown), PRM-701 (golden), and PRM-801 (white) grown under different nitrogen doses was determined by analyzing the growth, yield parameters and activities of nitrate reductase (NR), glutamine synthetase (GS), glutamate synthase; GOGAT, and glutamate dehydrogenase (GDH) at different developmental stages. High nitrogen use efficiency and nitrogen utilization efficiency were observed in PRM-1 genotype, whereas high nitrogen uptake efficiency was observed in PRM-801 genotype. At grain filling nitrogen uptake efficiency in PRM-1 negatively correlated with NR, GS, GOGAT activities whereas it was positively correlated in PRM-701 and PRM-801, however, GDH showed a negative correlation. Growth and yield parameters indicated that PRM-1 responds well at high nitrogen conditions while PRM-701 and PRM-801 respond well at normal and low nitrogen conditions respectively. The study indicates that PRM-1 is high nitrogen responsive and has high nitrogen use efficiency, whereas golden PRM-701 and white PRM-801 are low nitrogen responsive genotypes and have low nitrogen use efficiency. However, the crude grain protein content was higher in PRM-801 genotype followed by PRM-701 and PRM-1, indicating negative correlation of nitrogen use efficiency with source to sink relationship in terms of seed protein content.
PMCID: PMC3415157  PMID: 22919342
13.  Pediatric severe asthma is characterized by eosinophilia and remodeling without TH2 cytokines 
The pathology of pediatric severe therapy-resistant asthma (STRA) is little understood.
We hypothesized that STRA in children is characterized by airway eosinophilia and mast cell inflammation and is driven by the TH2 cytokines IL-4, IL-5, and IL-13.
Sixty-nine children (mean age, 11.8 years; interquartile range, 5.6-17.3 years; patients with STRA, n = 53; control subjects, n = 16) underwent fiberoptic bronchoscopy, bronchoalveolar lavage (BAL), and endobronchial biopsy. Airway inflammation, remodeling, and BAL fluid and biopsy specimen TH2 cytokines were quantified. Children with STRA also underwent symptom assessment (Asthma Control Test), spirometry, exhaled nitric oxide and induced sputum evaluation.
Children with STRA had significantly increased BAL fluid and biopsy specimen eosinophil counts compared with those found in control subjects (BAL fluid, P < .001; biopsy specimen, P < .01); within the STRA group, there was marked between-patient variability in eosinophilia. Submucosal mast cell, neutrophil, and lymphocyte counts were similar in both groups. Reticular basement membrane thickness and airway smooth muscle were increased in patients with STRA compared with those found in control subjects (P < .0001 and P < .001, respectively). There was no increase in BAL fluid IL-4, IL-5, or IL-13 levels in patients with STRA compared with control subjects, and these cytokines were rarely detected in induced sputum. Biopsy IL-5+ and IL-13+ cell counts were also not higher in patients with STRA compared with those seen in control subjects. The subgroup (n = 15) of children with STRA with detectable BAL fluid TH2 cytokines had significantly lower lung function than those with undetectable BAL fluid TH2 cytokines.
STRA in children was characterized by remodeling and variable airway eosinophil counts. However, unlike in adults, there was no neutrophilia, and despite the wide range in eosinophil counts, the TH2 mediators that are thought to drive allergic asthma were mostly absent.
PMCID: PMC3381727  PMID: 22385633
Pediatric asthma; eosinophilia; remodeling; severe therapy-resistant asthma; mediators
14.  Direct Analysis in Real Time by Mass Spectrometric Technique for Determining the Variation in Metabolite Profiles of Cinnamomum tamala Nees and Eberm Genotypes 
The Scientific World Journal  2012;2012:549265.
Cinnamomum tamala Nees & Eberm. is an important traditional medicinal plant, mentioned in various ancient literatures such as Ayurveda. Several of its medicinal properties have recently been proved. To characterize diversity in terms of metabolite profiles of Cinnamomum tamala Nees and Eberm genotypes, a newly emerging mass spectral ionization technique direct time in real time (DART) is very helpful. The DART ion source has been used to analyze an extremely wide range of phytochemicals present in leaves of Cinnamomum tamala. Ten genotypes were assessed for the presence of different phytochemicals. Phytochemical analysis showed the presence of mainly terpenes and phenols. These constituents vary in the different genotypes of Cinnamomum tamala. Principal component analysis has also been employed to analyze the DART data of these Cinnamomum genotypes. The result shows that the genotype of Cinnamomum tamala could be differentiated using DART MS data. The active components present in Cinnamomum tamala may be contributing significantly to high amount of antioxidant property of leaves and, in turn, conditional effects for diabetic patients.
PMCID: PMC3373138  PMID: 22701361
15.  Induction of MAP Kinase Homologues during Growth and Morphogenetic Development of Karnal Bunt (Tilletia indica) under the Influence of Host Factor(s) from Wheat Spikes 
The Scientific World Journal  2012;2012:539583.
Signaling pathways that activate different mitogen-activated protein kinases (MAPKs) in response to certain environmental conditions, play important role in mating type switching (Fus3) and pathogenicity (Pmk1) in many fungi. In order to determine the roles of such regulatory genes in Tilletia indica, the causal pathogen of Karnal bunt (KB) of wheat, semi-quantitative and quantitative RT-PCR was carried out to isolate and determine the expression of MAP kinase homologues during fungal growth and development under in vitro culture. Maximum expression of TiFus3 and TiPmk1 genes were observed at 14th and 21st days of culture and decreased thereafter. To investigate whether the fungus alters the expression levels of same kinases upon interaction with plants, cultures were treated with 1% of host factors (extracted from S-2 stage of wheat spikes). Such treatment induced the expression of MAPks in time dependent manner compared to the absence of host factors. These results suggest that host factor(s) provide certain signal(s) which activate TiFus3 and TiPmk1 during morphogenetic development of T. indica. The results also provides a clue about the role of host factors in enhancing the disease potential due to induction of MAP kinases involved in fungal development and pathogenecity.
PMCID: PMC3322648  PMID: 22547988
16.  Unusually large sialolith of Wharton's duct 
The formation of calcific concretions in the salivary duct or glands is a common disorder, especially in the submandibular glands. Most of the salivary calculi are small in size, in contrast to those that reach several centimeters, which are reported as megaliths or giant calculi in the literature. They may occur in any of the salivary gland ducts but are most common in Wharton's duct and the submandibular gland. This report presents clinical and radiographical sign of an unusually large sialolith. There was painless swelling on the floor of the edentulous mouth and patient was unaware of it. Radiographical examination revealed large irregular radio-opaque mass superimposed on right canine and premolar areas. This case report describes a patient presenting with an unusually large submandibular gland duct sialolith, the subsequent patient management, the aetiology, diagnosis and its treatment.
PMCID: PMC3591092  PMID: 23483770
Sialolith; submandibular salivary gland; wharton's duct
17.  Surgical management of ameloblastoma: Conservative or radical approach 
The ameloblastoma is a benign odontogenic tumor of epithelial origin that exhibits a locally aggressive behavior with a high level of recurrence, being believed theoretically to come from dental lamina remains, the enamel organ in development, epithelial cover of odontogenic cysts or from the cells of the basal layer of the oral mucosa. Especially larger, aggressive lesions require a more radical surgical approach resulting in large jaw defects. This paper discusses our experiences in the management of ameloblastoma tumor in 20 such patients.
Materials and Methods:
A review of 20 cases of ameloblastoma (6 in the maxillary and 14 in the mandibular region) is presented. The lesions were between 4 and 8 cm in diameter. The methods of treatment consisted of radical surgery (i.e., segmental resection) and conservative treatments (i.e., enucleation with bone curettage). Half the cases were treated conservatively and others surgically.
Enucleation with curettage was done in 10 cases, out of which six (60%) showed recurrence, whereas one (10%) case in the surgical group showed recurrence. Relatively higher tendencies of recurrence were observed in the cases treated conservatively. The aesthetic and functional outcomes were satisfying in all patients.
According to our opinion, radical surgical resection of ameloblastoma is the treatment of choice, followed by the reconstruction of the defects, allowing good functional and aesthetic outcome.
PMCID: PMC3304226  PMID: 22442605
Ameloblastoma; enucleation; resection
18.  A view from the other side of the table 
PMCID: PMC2083664  PMID: 17376954
Bone  2007;41(4):733-739.
Knowledge of the location of initial regions of failure within the vertebra — cortical shell, cortical endplates vs. trabecular bone, as well as anatomic location — may lead to improved understanding of the mechanisms of aging, disease and treatment. The overall objective of this study was to identify the location of the bone tissue at highest risk of initial failure within the vertebral body when subjected to compressive loading. Toward this end, micro-CT based 60-micron voxel-sized, linearly elastic, finite element models of a cohort of thirteen elderly (age range: 54–87 years, 75 ± 9 years) female whole vertebrae without posterior elements were virtually loaded in compression through a simulated disc. All bone tissue within each vertebra having either the maximum or minimum principal strain beyond its 90th percentile was defined as the tissue at highest risk of initial failure within that particular vertebral body. Our results showed that such high-risk tissue first occurred in the trabecular bone and that the largest proportion of the high-risk tissue also occurred in the trabecular bone. The amount of high-risk tissue was significantly greater in and adjacent to the cortical endplates than in the mid-transverse region. The amount of high-risk tissue in the cortical endplates was comparable to or greater than that in the cortical shell regardless of the assumed Poisson’s ratio of the simulated disc. Our results provide new insight into the micromechanics of failure of trabecular and cortical bone within the human vertebra, and taken together, suggest that during strenuous compressive loading of the vertebra, the tissue near and including the endplates is at the highest risk of initial failure.
PMCID: PMC2082110  PMID: 17643362
Vertebral body; Micromechanics; Initial failure; Trabecular bone; Cortical bone; Failure location

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