Bicameral rupture of aneurysm of sinus of valsalva

A rare case of bicameral rupture of aneurysm of noncoronary Sinus of Valsalva with severe aortic stenosis in an Indian female patient is reported. While closing the fistula from aorta, it is important to avoid any persistence of the fistula between the right atrium and ventricle beneath the patch.     

Transcorneal Permeation of l- and d-Aspartate Ester Prodrugs of Acyclovir: Delineation of Passive Diffusion Versus Transporter Involvement

PURPOSE: The aim of this study was to evaluate the contribution of amino acid transporters in the transcorneal permeation of the aspartate (Asp) ester acyclovir (ACV) prodrug. METHODS: Physicochemical characterization, solubility and stability of acyclovir L: -aspartate (L: -Asp-ACV) and acyclovir D: -aspartate (D: -Asp-ACV) were studied. Transcorneal permeability was evaluated across excised rabbit cornea. RESULTS: Solubility of L: -Asp-ACV and D: -Asp-ACV were about twofold higher than that of ACV. The prodrugs demonstrated greater stability under acidic conditions. Calculated pK(a) and logP values for both prodrugs were identical. Transcorneal permeability of L: -Asp-ACV [Formula: see text] was fourfold higher than D: -Asp-ACV [Formula: see text] and ACV [Formula: see text]. ACV generation during the transport process was minimal. L: -Asp-ACV transport was sodium and energy dependent but was not inhibited by glutamic acid. Addition of BCH, a specific B(0,+) and L amino acid transporter inhibitor, decreased transcorneal L: -Asp-ACV permeability to [Formula: see text]. L: -Asp-ACV and D: -Asp-ACV did not demonstrate significant difference in stability in ocular tissue homogenates. CONCLUSION: The results demonstrate that enhanced transport of L: -Asp-ACV is as a result of corneal transporter involvement (probably amino acid transporter B(0,+)) and not as a result of changes in physicochemical properties due to prodrug derivatization (permeability of D: -Asp-ACV and ACV were not significantly different).     

Impact of CYP3A5 and CYP3A4 gene polymorphisms on dose requirement of calcineurin inhibitors, cyclosporine and tacrolimus, in renal allograft recipients of North India

The present study investigated pharmacogenetic associations of common cytochrome P450 3A (CYP3A5 and CYP3A4) polymorphisms with dose requirements of calcineurin inhibitors, cyclosporine (CsA) and tacrolimus (Tac) in renal transplant recipients of North India. Two hundred twenty four patients on CsA and 73 patients on Tac-based immunosuppression regimen were genotyped for CYP3A5*3 (6986A>G) and CYP3A4*1B (-290A>G) and correlated with CsA/Tac dose requirement (mg/kg/day) and dose-adjusted CsA (C₂)/Tac (T ₀) blood levels (concentration/dose ratio) at 1 month and 3 months posttransplantation. The dose-adjusted levels were significantly lower in CYP3A5 expressers for CsA (p = 0.037; 3 months) and Tac (p < 0.001; 1 month and p < 0.001; 3 months) compared to the non-expressers, suggesting that for a given dose their CsA/Tac blood concentration is lower. The CYP3A5 non-expresser genotype was associated with reduced risk for allograft rejection (HR-0.18, 95% CI 0.03–0.99). No influence of CYP3A4*1B on CsA/Tac pharmacokinetics was observed. CYP3A5 expressers were associated with significantly lower dose-adjusted CsA/Tac concentrations and higher allograft rejection episodes in patients on Tac therapy.     

Chromium picolinate induced apoptosis of lymphocytes and the signaling mechanisms thereof

Cr(III)(picolinate)₃ [Cr(III)(pic)₃] is currently used as a nutritional supplement and for treating Type-2 diabetes. The effect of Cr(III)(pic)₃ uptake in peripheral blood lymphocytes is investigated in this study. From the cytotoxicity data, DNA fragmentation pattern, Annexin V staining, TUNEL positivity and the ultrastructural characteristics such as chromatin condensation and formation of apoptotic bodies, it is clear that Cr(III)(pic)₃ induces a concentration dependent apoptosis. It is shown that reactive oxygen species (ROS) produced by treatment with Cr(III)(pic)₃ leads to apoptosis, since we find that pretreatment with N-acetyl cysteine inhibits the process. Using Western blotting technique and fluorescence measurements, the downstream signaling molecules have also been identified. Cr(III)(pic)₃ treatment leads to collapse of the mitochondrial membrane potential, Bax expression, increase in cytosolic cytochrome c content and active caspase-3 and DNA fragmentation and all these manifestations are reduced by pretreating the lymphocytes with N-acetyl cysteine. Thus, it is shown that Cr(III)(pic)₃ is cytotoxic to lymphocytes with ROS and mitochondrial events playing a role in bringing about apoptosis.     

Effect of Ocimum sanctum, ascorbic acid, and verapamil on macrophage function and oxidative stress in mice exposed to cocaine

Objective:

To investigate the effect of Ocimum sanctum, ascorbic acid, and verapamil on macrophage function and oxidative stress in experimental animals exposed to cocaine.

Materials and Methods:

Mice were used in this study and were divided randomly into different groups of six animals each. They were either treated with intraperitoneal injection of saline or cocaine hydrochloride or an oral feeding of oil of Ocimum sanctum, ascorbic acid or verapamil, or both (ascorbic acid and verapamil), and were evaluated for a respiratory burst of macrophages, superoxide and nitric oxide (NO) production, estimation of TNF-α in the serum and supernatant of cultured macrophages, estimation of lipid peroxidation (malondialdehyde- MDA) in the serum, and superoxide dismutase activity in the erythrocytes.

Results:

Unstimulated respiratory burst as well as superoxide production was enhanced on treatment with cocaine and all the three drugs were found to attenuate this enhancement. The bactericidal capacity of macrophages decreased significantly on chronic cocaine exposure, as it was associated with decreased respiratory burst and superoxide production. There was a significant decrease in NO production by macrophages on chronic cocaine exposure and all the test drugs were found to restore nitrite formation to a normal level. There was an increase in the malonylodialdehyde (MDA) level and decrease in the superoxide dismutase level on chronic cocaine exposure, and all the three drugs effectively decreased the MDA level and increased superoxide dismutase level. There was an increase in serum TNF-α on chronic cocaine exposure, which was decreased significantly by ascorbic acid and verapamil.

Conclusion:

O. sanctum, ascorbic acid, and verapamil were equally effective in improving the macrophage function and reducing oxidative stress. These findings suggested that O. sanctum, ascorbic acid, and verapamil attenuated acute and chronic cocaine-mediated effects.     

Multiplex real-time PCR assay for simultaneous detection of Acanthamoeba spp., Balamuthia mandrillaris, and Naegleria fowleri

Infections caused by Naegleria fowleri, Acanthamoeba spp., and Balamuthia mandrillaris occur throughout the world and pose many diagnostic challenges. To date, at least 440 cases of severe central nervous system infections caused by these amebas have been documented worldwide. Rapid and specific identification of these free-living amebas in clinical samples is of crucial importance for efficient case management. We have developed a triplex real-time TaqMan PCR assay that can simultaneously identify Acanthamoeba spp., B. mandrillaris, and N. fowleri in the same PCR vessel. The assay was validated with 22 well-characterized amebic strains harvested from cultures and nine clinical specimens that were previously characterized by in vitro culture and/or immunofluorescence assay. The triplex assay demonstrated high specificity and a rapid test completion time of less than 5 h from the reception of the specimen in the laboratory. This assay was able to detect one single ameba per sample analyzed, as determined with cerebrospinal fluid spiked with diluted cultured amebas. This assay could become useful for fast laboratory diagnostic assessment of amebic infections (caused by free-living amebas) in laboratories with adequate infrastructure to perform real-time PCR testing.     

Buerger's disease: clinical and histomorphological study

There is an extremely high prevalence of Thromboangiitis Obliterans (TAO) or Buerger's disease (BD) in India among people of low socioeconomic class who smoke beedies (homemade cigarettes with raw tobacco). The aim of this study was to study the clinical and histo-morphological aspects of Buerger's disease with relevance to age at presentation in the local population. The study comprised of 25 cases (all were men and were smokers) of clinically diagnosed BD based on Shionoya's criteria. The mean age was 47 years. The specimens consisted of 21 biopsies, 2 end-arterectomies and 2 amputations. Formalin fixed, routinely processed, paraffin embedded tissue sections were stained with Haematoxylin and Eosin (H and E) and Verhoeff's elastic stain. They had claudication pain either in the ankle (5) or in the calf (2) or both (13). 24 had infrapopliteal disease and 9 showed upper limb involvement. 21 showed migratory thrombophlebitis also. Histomorphological presentation included the following features: Luminal thrombosis (14), fresh thrombosis (4), chronic inflammation in the vessel wall (10), within the thrombus (1) and around perivascular channels and nerve bundles (4). Internal elastic lamina showed reduplication in 13, undulation in 9 and fragmentation in 9 cases. Media of the vessel showed the following features: fibrosis (9), hypertrophy (9) and calcification (5) Adventitial haemorrhage, cholesterol clefts and atherosclerotic plaque formation were the other changes seen. In our study the following histopathological features were consistently seen. Thrombus (with or without recanalisation), inflammatory cell infiltrate (within the thrombus wall or periadventitial tissue), subintimal and medial fibrosis and changes in internal elastic lamina. These features were also highlighted in other studies. However in our study, medial hypertrophy and calcification were observed as additional features.     

Tissue Inhibitor of Metalloproteinases 3 Regulates Resolution of Inflammation following Acute Lung Injury

Tissue inhibitor of metalloproteinases 3 (TIMP3) inhibits not only matrix metalloproteinases but also a disintegrin and metalloproteinase domain family members and thus contributes to controlling diverse processes mediated by proteolysis. We used Timp3^−/− mice to assess the role of this inhibitor in acute lung injury. After bleomycin-induced injury, inflammation, as indicated by the influx of neutrophils in bronchoalveolar lavage (BAL), peaked at 7 days post-injury in the wild-type mice and began to wane thereafter; however, in Timp3^−/− mice, inflammation persisted up to 28 days. Furthermore, although the level of chemokines in BAL and lung homogenate was similar in both genotypes, BAL from Timp3^−/− mice 7, 14, and 28 days post-injury had increased neutrophil chemotactic activity compared with wild-type BAL. At day 14, a higher percentage of apoptotic neutrophils were present in wild-type mice compared with Timp3^−/− mice, further suggesting that TIMP3 constrains continued neutrophil influx. In addition, total matrix metalloproteinase activity was increased in lungs from Timp3^−/− mice, and treatment of mice with a synthetic inhibitor of metalloproteinases rescued the enhanced neutrophilia phenotype. These data demonstrate that TIMP3 regulates neutrophil influx in the lung following injury through its ability to inhibit metalloproteinase activity and indicates that TIMP3 functions to promote the resolution of inflammation in the lung.Acute lung injury, which often results from severe pulmonary infection or trauma and can progress to acute respiratory distress syndrome, is characterized by profound inflammation and tissue injury.¹ Recovery from acute lung injury requires the proper regulation of wound repair, inflammation, and the deposition and remodeling of extracellular matrix.² Many of these processes are mediated by matrix metalloproteinases (MMPs), which in turn are balanced by their natural inhibitors, the tissue inhibitors of metalloproteinases (TIMPs).^2–4TIMP3, which is expressed and stored in the lung matrix, is unique from other TIMPs in that in addition to inhibiting MMPs, it is also an efficient inhibitor of multiple members of the a disintegrin and metalloproteinase (ADAM) domain family, including ADAM17 or tumor necrosis factor α (TNFα)-converting enzyme.³ The importance of TIMP3 in lung homeostasis is highlighted by the development of enlarged alveolar spaces in aging TIMP3-null (Timp3^−/−) mice.⁵ Additionally, adult Timp3^−/− mice have altered inflammatory responses in various tissues. For example, following partial hepatectomy or systemic LPS exposure, circulating levels of active TNFα are increased in Timp3^−/− mice compared with wild-type mice.^6,7 Furthermore, shedding of TNFα is increased in macrophages isolated from Timp3^−/− mice,⁷ suggesting that TIMP3 controls the proinflammatory activation of macrophages in a cell-autonomous manner.Several members of both the MMP and TIMP families have been demonstrated to control various processes in response to lung injury. For example, our group and others demonstrated that mice lacking matrilysin (MMP7) are protected from bleomycin-induced lung injury due to impaired neutrophil migration into the air space during the acute phase and to a later modest reduction in fibrosis.^8,9 Mice lacking TIMP1 reveal phenotypes in response to bleomycin injury that are nearly opposite to those seen in Mmp7^−/− mice, such as enhanced neutrophil influx into the alveolar space,^10,11 likely due to its ability to inhibit MMP7.^10,11 Overexpression of TIMP1 in the lung epithelium, however, does not protect against the deleterious effects of bleomycin,¹² suggesting that the endogenous levels of this inhibitor are well tuned to govern the activity of proteinases it targets.Recently, microarray analysis of BALB/c (which are resistant to bleomycin-induced lung injury) and C57BL/6 mice (which are susceptible to bleomycin-induced lung injury) revealed that Timp3 expression is reduced in C57BL/6 mice—compared with its elevated expression in BALB/c—during the initial, damaging inflammatory response.¹³ These data predict that TIMP3 functions to moderate the host response to tissue injury. Indeed, we report that whereas TIMP3 does seem to affect early response to injury, it is critical for proper resolution of inflammation and fibrosis at later stages. These findings suggest that this natural inhibitor functions to block the activity of metalloproteinases (both MMP and ADAM) mediating proinflammatory and fibrotic pathways.