Structural and therapeutic properties of salicylic acid-solubilized Pluronic solutions and hydrogels.

Salicylic acid (SA) finds extensive applications in the treatment of rheumatic and skin diseases because of its analgesic, anti-inflammatory and exfoliating properties. As it is lipophilic in nature, there is a need for appropriate delivery systems to harness these properties for different applications. Herein, we examined the suitability of Pluronic P123/F127 micellar systems as delivery media by investigating the structural, flow and antimicrobial properties of P123/F127-SA solutions and hydrogels using DLS, SANS, rheological and zone inhibition measurement techniques. SA modulates the aggregation characteristics of these surfactant systems and brings about spherical-to-worm-like micelle-to-vesicular structural transitions in the hydrophobic Pluronic P123 system, a spherical-to-worm-like micellar transition in the mixed P123/F127 system and an onset of inter-micellar attraction in the hydrophilic Pluronic F127 system. SA-solubilized systems of both hydrophobic and hydrophilic Pluronics inhibit the growth of Gram-positive and Gram-negative bacteria with comparable MIC values. This suggests that the interaction of SA molecules with the bacterial cell membrane remains unobstructed upon encapsulation in Pluronic micelles. F127 hydrogel-based SA formulations with rheological properties suitable for topical applications and up to 15% SA loading were prepared. These will be useful SA ointments as F127 is an FDA-approved excipient for topical drug delivery applications. The results indicate that Pluronics remain effective as delivery agents for SA and exhibit interesting structural polymorphism upon its solubilization.

Thrombospondin-1 triggers macrophage IL-10 production and promotes resolution of experimental lung injury.

Mononuclear phagocyte recognition of apoptotic cells triggering suppressive cytokine signaling is a key event in inflammation resolution from injury. Mice deficient in thrombospondin (TSP)-1 (thbs1⁻/⁻), an extracellular matrix glycoprotein that bridges cell-cell interactions, are prone to lipopolysaccharide-induced lung injury and show defective macrophage interleukin (IL)-10 production during the resolution phase of inflammation. Reconstitution of IL-10 rescues thbs1⁻/⁻ mice from persistent neutrophilic lung inflammation and injury and thbs1⁻/⁻ alveolar macrophages show defective IL-10 production following intratracheal instillation of apoptotic neutrophils despite intact efferocytosis. Following co-culture with apoptotic neutrophils, thbs1⁻/⁻ macrophages show a selective defect in IL-10 production, whereas prostaglandin E2 and transforming growth factor beta 1 responses remain intact. Full macrophage IL-10 responses require the engagement of TSP-1 structural repeat 2 domain and the macrophage scavenger receptor CD36 LIMP-II Emp sequence homology (CLESH) domain in vitro. Although TSP-1 is not essential for macrophage engulfment of apoptotic neutrophils in vivo, TSP-1 aids in the curtailment of inflammatory responses during the resolution phase of injury in the lungs by providing a means by which apoptotic cells are recognized and trigger optimal IL-10 production by macrophages.

Phacomatosis cesioflammea with Klippel Trenaunay syndrome: A rare association.

A 30-year-old Indian male presented with bilateral Nevus of Ota, extensive nevus flammeus over the trunk and left lower limb with soft tissue hypertrophy and varicosities affecting the left lower limb. He was otherwise in good general health. A diagnosis of Phacomatosis cesioflammea or Phacomatosis pigmentovasularis Type II with Klippel Trenaunay syndrome was made. The case is being reported on account of its rarity.

Conjunctival impression cytology in computer users.

INTRODUCTION: It is known that the computer users develop the features of dry eye. OBJECTIVE: To study the cytological changes in the conjunctiva using conjunctival impression cytology in computer users and a control group. MATERIALS AND METHODS: Fifteen eyes of computer users who had used computers for more than one year and ten eyes of an age-and-sex matched control group (those who had not used computers) were studied by conjunctival impression cytology. RESULTS: Conjunctival impression cytology (CIC) results in the control group were of stage 0 and stage I while the computer user group showed CIC results between stages II to stage IV. Among the computer users, the majority ( > 90 %) showed stage III and stage IV changes. CONCLUSION: We found that those who used computers daily for long hours developed more CIC changes than those who worked at the computer for a shorter daily duration.

STAT1-regulated lung MDSC-like cells produce IL-10 and efferocytose apoptotic neutrophils with relevance in resolution of bacterial pneumonia.

Bacterial pneumonia remains a significant burden worldwide. Although an inflammatory response in the lung is required to fight the causative agent, persistent tissue-resident neutrophils in non-resolving pneumonia can induce collateral tissue damage and precipitate acute lung injury. However, little is known about mechanisms orchestrated in the lung tissue that remove apoptotic neutrophils to restore tissue homeostasis. In mice infected with Klebsiella pneumoniae, a bacterium commonly associated with hospital-acquired pneumonia, we show that interleukin (IL)-10 is essential for resolution of lung inflammation and recovery of mice after infection. Although IL-10(-/-) mice cleared bacteria, they displayed increased morbidity with progressive weight loss and persistent lung inflammation in the later phase after infection. A source of tissue IL-10 was found to be resident CD11b(+)Gr1(int)F4/80(+) cells resembling myeloid-derived suppressor cells (MDSCs) that appeared with a delayed kinetics after infection. These cells efficiently efferocytosed apoptotic neutrophils, which was aided by IL-10. The lung neutrophil burden was attenuated in infected signal transducer and activator of transcription 1 (STAT1)(-/-) mice with concomitant increase in the frequency of the MDSC-like cells and lung IL-10 levels. Thus, inhibiting STAT1 in combination with antibiotics may be a novel therapeutic strategy to address inefficient resolution of bacterial pneumonia.

Terminal 4q deletion syndrome.

Terminal deletion of the long arm of chromosome 4, (4q) is a rare event. It is characterized by spectral phenotypic manifestations, depending upon the site and quantity of chromatin lost. The chromosomal loss which span 4 (q31-q35) segment often manifests as craniofacial anomalies, mental retardation with ocular, cardiac, genitourinary defects and pelvic/limb dysmorphism. These abnormalities are usually unilateral. We report a female child (46, XX), aged 11 months, born to nonconsanguineous parents, bearing chromosomal deletion of 4 (q31.2-35.2) segment, which has manifested as craniofacial hypoplasia of left side of face, ipsilateral ptosis, erythroderma and bilateral thumb anomalies.

Oral propranolol--efficacy and comparison of two doses for peri-operative anxiolysis.

Patients undergoing surgery are having high levels of anxiety and stress. Though not life threatening it adds an unnecessary financial burden to the hospital. We assessed the anxiolytic effects of oral propranolol 20 mg and 40 mg when given as premedication. This is a double blind, randomised, prospective clinical study involving 60 healthy patients (ASA I and II) undergoing minor elective surgery. Subjects in control group without any anxiolytic premedication (group I, n = 20) were compared with those receiving oral propranolol 20 mg (group II, n = 20) or 40 mg (group III, n = 20) with sips of water 2-hour prior to surgery. Anxiety level was assessed using 4-point anxiolysis score (1--tearful, 2--anxious but easily reassured, 3--calm, 4--asleep) in the holding area, after entering operating room, immediate postoperative and 2 hours after surgery. Fluctuation in pulse rate and BP was recorded. Statistical data was analysed using one-way ANOVA with posthoc test.Value of p < 0.05 was taken as significant. Twenty subjects in each group were required as calculated from reference study with difference of up to 30%, type I error of 0.5 and power of 80%. Anxiolysis score in operating room (group I--1.40 +/- 0.48, group II--1.95 +/- 0.58, group III--1.90 +/- 0.53) and immediate postoperative period (group I--1.25 +/- 0.43, group II--1.90 +0.53, group III--2.10 +/- 0.29) were significantly improved (p < 0.05) in groups II and III compared with control group. Variations in systolic BP and pulse rate were less in test groups (p < 0.05). No statistically significant difference was found after Intergroup comparison of test groups. Bradycardia (25%) and hypotension (10%) were more with 40 mg propranolol. Both 20 mg and 40 mg doses of propranolol are effective for pre-operative anxiolysis but 20 mg dose gives significant reduction in anxiety with minimal side-effects. Thus 20 mg propranolol premedication for reducing peri-operative anxiety and for cardiovascular stability is recommended.

Kaposi's sarcoma as a presenting manifestation of HIV.

Kaposi's sarcoma is a multi-focal vascular tumor involving skin and the other organs. HIV associated Kaposi's sarcoma is one of the AIDS defining condition. It is rarely reported from India. We report a 40-year-old heterosexual married male with widespread cutaneous lesions of Kaposi's sarcoma without any oral lesions or systemic association as a presenting manifestation of HIV.

Regulatory T cells in many flavors control asthma.

That regulatory T cells (Tregs) have a crucial role in controlling allergic diseases such as asthma is now undisputed. The cytokines most commonly implicated in Treg-mediated suppression of allergic asthma are transforming growth factor-beta (TGF-beta) and interleukin (IL)-10). In addition to naturally occurring Tregs, adaptive Tregs, induced in response to foreign antigens, have been shown in recent studies. The concept of inducible/adaptive Tregs (iTregs) has considerable significance in preventing asthma if generated early enough in life. This is because cytokines such as IL-4 and IL-6 inhibit Foxp3 induction in naive CD4+ T cells and therefore de novo generation of Tregs can be expected to be less efficient when it is concomitant with effector cell development in response to an allergen. However, if iTregs can be induced, the process of infectious tolerance would facilitate expansion of the iTreg pool as suggested in the recent literature. It is tempting to speculate that there is a window of opportunity in early life in the context of a relatively immature immune system that is permissive for the generation of iTregs specific to a spectrum of allergens that would regulate asthma for lifelong. The focus of this review is the relevance of nTregs and iTregs in controlling asthma from early life into adulthood, the mechanisms underlying Treg function, and the prospects for using our current concepts to harness the full potential of Tregs to limit disease development and progression.

Thiol metabolism play significant role during cadmium detoxification by Ceratophyllum demersum L.

In the present study, the level of thiols and activity of related enzymes were investigated in coontail (Ceratophyllum demersum L.) plants to analyze their role in combating the stress caused upon exposure to cadmium (Cd; 0-10 microM) for a duration up to 7d. Plants showed the maximum accumulation of 1293 microg Cd g(-1)dw after 7d at 10 microM. Significant increases in the level of total non-protein thiols (NP-SH) including phytochelatins (PCs) as well as upstream metabolites of the PC biosynthetic pathway, cysteine and glutathione (GSH) were observed. In addition, significant increases in the activities of cysteine synthase (CS), glutathione-S-transferase (GST), glutathione reductase (GR), as well as in vitro activation of phytochelatin synthase (PCS), were noticed in response to Cd. In conclusion, under Cd stress, plants adapted to a new metabolic equilibrium of thiols through coordinated synthesis and consumption to combat Cd toxicity and to accumulate it.

Histopathological manifestations in kidney of Clarias batrachus induced by experimental Procamallanus infection.

Kidney of Clarias batrachus infected with Procamallanus showed varying degrees of histopathological alterations on 15, 30, 45 and 60 days post-infection. The infected kidney showed variable sized glomeruli, cloudy swelling in tubules, vacuolar/atrophic degeneration, fibrosis, mild degenerative changes in distal convoluted tubules, enlarged Bowmen's capsule, necrotic changes as well as increased granulation and hyperplasia in proximal convoluted tubules after 15 days. After 30 days of infection, the changes were rupture of Bowmen's capsule wall, degenerative changes, edema, necrosis, pyknosis, karyorrhexis and karyolysis in proximal and distal convoluted tubules, fibrosis, cloudy swelling and inflammatory lymphocytes, proliferation and shrinkage in glomeruli, and vacuolization in proximal convoluted tubules as well as cloudy swelling. After 45 days, the infected kidney showed cloudy swelling in glomeruli as well as variation in their size, infiltration of RBCs in intralobular vein and necrosis in proximal convoluted tubules, cloudy swelling in interstitium, vacuolization in the epithelial lining cells, necrosis in haemopoietic tissue and inflammatory edema. After 60 days post-infection, the changes were rupture of intralobular vein, cloudy swelling, necrosis in few proximal convoluted tubules, atrophy and shrinkage in glomeruli, distinct inflammatory edema, pyknosis, karyorrhexis and karyolysis, aggregation of lymphocytes and dilation in blood vessels.

Flowcytometric evidence of platelet activation in patients on aspirin following myocardial infarction.

BACKGROUND: Following a myocardial infarction, patients are usually started on long term antiplatelet therapy with aspirin in a dose of 80-150 mg/day. However, there are no quick and easy methods to assess the efficacy of the antiplatelet activity of aspirin. METHODS: We studied 60 consecutive patients (men, < 40 years of age) 8-10 weeks after they had had acute myocardial infarction. These patients were receiving 100 mg aspirin daily orally with or without b-blockers. We measured P-selectin expression and fibrinogen binding by flowcytometry at least 3 times over a period of 2 years in all the patients. We also studied 100 age- and sex-matched controls. RESULTS: Of the 60 patients, 30 (50%) showed both increased P-selectin and fibrinogen binding by platelets, suggesting platelet activation. Fourteen other patients had increased fibrinogen binding but normal P-selectin expression. Sixteen patients and all the controls had normal results of both tests. CONCLUSION: Our data show evidence of platelet activation in at least 50% of patients receiving 100 mg of aspirin daily. Flowcytometry for P-selectin expression and fibrinogen binding to platelets can be used to monitor antiplatelet therapy with aspirin following acute myocardial infarction.

Evaluation of markers of endothelial damage in cases of young myocardial infarction.

The pathogenesis of arterial thrombotic disease involves multiple genetic and environmental factors related to atherosclerosis and thrombosis. The endothelium is a monolayer of polygonal cells that extend continuously over the luminal surface of the entire vasculature. Injury to the endothelium leads to dysfunction. The causes of injury include lipids, immune complexes, microorganisms, smoking, hypertension, aging, diabetes mellitus and trauma. Studies have been done to evaluate the role of different adhesion molecules on the endothelial membrane in the pathogenesis of atherosclerosis. These molecules are intercellular adhesion molecule type-1 (ICAM-1), vascular cell adhesion molecule type-1 (VCAM-1), platelet/endothelial cell adhesion molecule-1 (PECAM-1), soluble P-selectin (sP-selectin) and soluble E-selectin (sE-selectin). One-hundred and twenty patients of myocardial infarction (age below 40 years) were recruited from the out-patients department of Department of Cardiology, KEM Hospital, Mumbai. All the patients were recruited 8-10 weeks after stabilization after MI. We estimated the levels of sP-selectin, sE-selectin, sPECAM-1 and serum homocysteine. Healthy age and sex-matched controls and family controls were also recruited in the present study. The levels of sP-selectin, sE-selectin and sPECAM-1 did not differ significantly in cases as compared to controls (p>0.05). Hyperhomocysteinemia was significantly associated with MI in comparison with controls (p<0.001) with an odds ratio of 6.26 (95% confidence limits 3.11-12.76). Folic acid was able to correct hyperhomocysteinemia in a large majority of the cases. Although the levels of sP-selectin, sE-selectin and sPECAM-1 decreased after folic acid therapy, it was only sE-selectin which was significantly reduced (p<0.05). Thus, folic acid had a dual effect in that it reduced hyperhomocysteinemia and sE-selectin which showed a significant reduction on folate supplementation for 15 days.

Thrombophilia in coronary artery disease: a double jeopardy.

Thrombophilia can be defined as an increased risk of thrombosis. The central event to the pathogenesis of any thrombotic episode is the perturbation of haemostasis, the cause of which may be genetic or environmental. The clinical manifestations of the chronic development of coronary artery atheroma are angina and acute myocardial infarction. In recent years literature is emerging on the role of different factors of blood coagulation in arterial thrombosis. Different coagulation factors, natural anticoagulants, platelet antigens and other factors such as homocysteine, lipoprotein (a), have been studied as risk factors for coronary artery disease (CAD). The results of many of these studies are contradictory. In India, there is an alarming rise in the number of young patients with myocardial infarction (MI) and an interesting feature is that a large majority of these patients lack the conventional risk factors. There have been scattered studies on the thrombophilia status among Indians. The management of thrombophilia can be done by a regimen of different drugs which has been evaluated in different clinical trials. Since the cost of thrombophilia investigations is quite phenomenal for a developing country like India, the selection of these investigations assumes an utmost importance.

Platelet function tests using platelet aggregometry: need for repetition of the test for diagnosis of defective platelet function.

Four hundred and ninety seven patients were referred to our center for platelet aggregation studies because of spontaneous mucocutaneous bleeds. All these patients had normal complete blood count, platelet count and peripheral smears except in ten patients of Bernard Soulier Syndrome where platelet count was marginally reduced in the presence of giant platelets. Two hundred and eighty patients were found to have normal platelet aggregation to ADP, collagen, ristocetin and arachidonic acid. Out of the remaining 217 patients, 62 patients were diagnosed to have Glanzmanns thrombasthenia, 10 Bernard Soulier Syndrome, 6 storage pool deficiencies, 7 cyclooxygenase deficiencies and 72 von Willebrand disease. In all the patients with GT and BSS, diagnosis was confirmed with flow cytometry using multiple monoclonal antibodies to GPIIb-IIIa and GPIb-IX. There were sixty patients where initial platelet aggregation studies showed reduced (<30%) aggregation to either ADP, collagen, ristocetin or arachidonic acid in its various combination, however in 12 such patients (20%) the platelet aggregation studies were normal on repetition. All our platelet aggregation studies were done only after assuring that the patient is not taking any medicine for at least 7-10 days which may affect the platelet function tests. The present study shows that single atypically abnormal platelet aggregation studies should always be repeated. Finally in 48/217 patients (22%) some aggregation abnormality to one or more of the agonists persisted, although we could not categorize these patients into any clear-cut platelet disorder. None of these 48 patients platelet associated immunoglobulin was increased by flow cytometry. It is possible that large number of patients from that disorder will finally prove to be some form of platelet secretory defect. In north India similar group of defect in a large number of patients have been reported as isolated PF3 abnormality or thrombasthenic thrombopathy.

Hepatic involvement and hepatitis B surface antigen (Hbs Ag) in leprosy.

Hepatic involvement and hepatitis B surface antigenemia was studied in 80 leprosy patients and results were compared with 50 normal healthy controls. HbsAg was detected in 7.54% of lepromatous leprosy patients as compared to 2% of the normal healthy controls. There was a decrease in albumin and increase in globulin levels with significant decrease in A: G ratio. SGPT levels were significantly raised in lepromatous leprosy patients. Histopathological changes were present in 57.1% of lepromatous leprosy and 23.8% of tuberculoid leprosy patients.