Antitumor and anticarcinogenic actions of Cpd 5: a new class of protein phosphatase inhibitor

Dual specificity phosphatases (DSP) play an important role in control of the cell cycle and signal transduction. We have synthesized a new class of DSP inhibitors. Cpd 5 or [2-(2-mercaptoethanol)-3-methyl-1,4-naphthoquinone] is one of the most potent of these. It inhibits DSPs of cells in culture and induces tyrosine phosphorylation of various DSP substrates, including the Cdc25 target Cdks and it potently inhibits cell growth. In this study, we have evaluated Cpd 5 in vivo for its antitumor and growth inhibitory activity on carcinogen-altered foci. Cpd 5 inhibited growth of the transplantable rat hepatoma cell line JM-1 in vitro, with concomitant phosphorylation of the mitogen-activated protein kinase ERK1/2 but not JNK1/2 or p38. This ERK phosphorylation was associated with growth inhibition, as the ERK phosphorylation inhibitor PD098059 antagonized both ERK phosphorylation and growth inhibition. JM-1 cell lysates were found to contain ERK1/2-specific phosphatase(s) that could be inhibited by Cpd 5 and which are thought to be its major targets. Cpd 5 caused significant inhibition of both intrahepatic and subcutaneous (s.c.) growth of transplanted JM-1 cells in male Fischer F344 rats. The treatment was equally effective whether Cpd 5 was administered either as a single, acute dose or chronically as several lower doses. However, toxicity was much lower with chronic treatment. As in JM-1 cells in vitro, ERK1/2 was phosphorylated when rats in vivo were treated with Cpd 5 and tumor growth inhibition in vivo also was antagonized by treating rats with the ERK1/2 phosphorylation inhibitor PD098059. A single dose of Cpd 5 also inhibited the formation of glutathione S-transferase-pi enzyme-altered cells induced by the hepatocarcinogen N-nitrosodiethylamine. This is the first report of the in vivo activity and growth inhibitory mechanism of a novel class of K vitamin growth inhibitors that have potent tyrosine phosphatase activity.     

Utility of transient elastography (fibroscan) and impact of bariatric surgery on nonalcoholic fatty liver disease (NAFLD) in morbidly obese patients

Background

Controlled attenuation parameter (CAP) is a novel, noninvasive technique for assessing hepatic steatosis. However, its role in morbidly obese individuals is unclear. The effect of bariatric surgery on inflammation and fibrosis needs to be explored.

Detail microscopic analysis of deep fascia of lower limb and its surgical implication

Background:

The knowledge regarding the structural details of deep fascia remains inadequate. It was described to be relatively avascular having predominantly protective function. Anatomical and surgical studies revealed that it had associated vascular arcade and hence incorporated it to ascertain additional vascularity to the flaps. However, not much importance has been directed towards the detailed study of the various constituents of deep fascia in order to explain its physiological and clinical implications. Therefore, this study was undertaken to unveil these details.

Materials and Methods:

Fifty fresh specimens of human deep fascia overlying the gastrocnemius muscle were analyzed regarding the (i) vasculature, (ii) matrix, and (iii) other structural elements. The deep fascia was procured in three forms; (a) both the layers, (b) superficial layer, and (c) deep layer. Detail study was conducted by light, confocal, and electron microscopy.

Results:

Under light microscopy, blood vessels including capillaries were seen associated with both the layers. Perforators traversing the intra-fascial plane could be visualized. Confocal microscope optical sections showed well-organized bright fluorescent collagen fibers and nuclei of various cells. Electron microscopic evaluation revealed many interesting constituents which are relatively unknown to the anatomist and clinicians. There were arterioles, capillaries, venules, lymphatics, nerves, mast cells, and myofibroblasts apart from collagen and elastic fibers.

Conclusion:

The detail structural analysis of deep fascia provided the clue to its rich vascularity and other structural constituents. They all contribute to enhance the vascularity and maintenance of the physiological functions of fasciocutaneous, adipofascial, and fascial flaps, frequently used for reconstructions. Thus, incorporation of deep fascia in the flaps during reconstruction is highly beneficial for ensuring optimal vascularity.     

Effect of surgical traumas on microcirculation

Background:

Adequate microcirculation in different tissues maintains the physiological function and heals surgical wounds. In any surgical procedure, the commonly used instruments are cautery, tissue forceps, and clamps. The fact that their inappropriate use produces an adverse effect on microcirculation is often not realized. By this study, we could demonstrate live, the effect of these surgical traumas.

Methods:

The study was conducted on the inferiorly based fasciocutaneous flap with a fascial extension in patients with a distal leg defect. The extended fascial flap was mounted on a glass slide and observed for live microcirculation under ×160 magnification. Three methods were used: (a) cautery in low power, (b) microcrushing forceps to crush the vessels, and (c) noncrushing clamps at the base of the fascial flap.

Results:

It was observed that the vessels are well protected within the deep fascia. Once the fascia was pierced the current damaged the vessel wall. As the wattage was increased, it caused charring of the tissue and multiple vessels ultimately leading to cessation of blood flow. Once the vessel wall was crushed by forceps, blood extravasated in a variable intensity depending upon the size of the vessel. Clamping led to gradual slowing of blood flow with microclot formation. In certain vessels, there was discontinuity in the blood column and ultimately the blood flow stopped.

Conclusion:

This study showed live demonstration of the effect of surgical traumas on microcirculation. It should guide the surgeons to select the use of appropriate instruments which will cause minimal damage to vascularity and thereby lead to a better surgical outcome.     

Left atrial extension of lung malignancy with ECG changes resembling STEMI

Lung malignancy extending into left atrium is seen very infrequently. We had a patient with a fast growing symptomatic lung mass and electrocardiogram showing persistent coving ST elevation without any biomarker change. Transthoracic echocardiography showed a large left atrial mass which was fixed to the free walls and extended into the appendage. There was also a large lung mass that was compressing the heart from its lateral aspect. CT-scan of chest corroborated the lung mass & CT-guided FNAC showed small cell carcinoma.     

Activin A promotes multiple myeloma-induced osteolysis and is a promising target for myeloma bone disease

Understanding the pathogenesis of cancer-related bone disease is crucial to the discovery of new therapies. Here we identify activin A, a TGF-β family member, as a therapeutically amenable target exploited by multiple myeloma (MM) to alter its microenvironmental niche favoring osteolysis. Increased bone marrow plasma activin A levels were found in MM patients with osteolytic disease. MM cell engagement of marrow stromal cells enhanced activin A secretion via adhesion-mediated JNK activation. Activin A, in turn, inhibited osteoblast differentiation via SMAD2-dependent distal-less homeobox–5 down-regulation. Targeting activin A by a soluble decoy receptor reversed osteoblast inhibition, ameliorated MM bone disease, and inhibited tumor growth in an in vivo humanized MM model, setting the stage for testing in human clinical trials.     

Role of apoptotic regulators in human epithelial ovarian cancer

We assessed molecular markers such as BRCA1, K-ras, p53, Bcl-2, Bcl-XL, Survivin and telomerase activity in untreated ovarian cancer tissue samples, ascitic cells and normal ovarian tissues and gathered insights into their correlation with each other and also with apoptotic index. The expression of these proteins was analyzed by Western blotting and immunohistochemistry. Apoptotic index was determined by TUNEL assay and telomerase activity was measured by PCR-ELISA kit. p53, Bcl-2, Bcl-XL, K-ras and Survivin were found to be over expressed in tumors and ascitic cells as compared to normal controls whereas there was no significant difference in expression of BRCA1. A significantly higher telomerase activity and lower apoptotic index in tumors as compared to controls was observed. p53 positively correlated with Bcl-2, Bcl-XL, K-ras and Survivin expression and also clinical stage of the disease. A positive correlation between Survivin and Bcl-2, Bcl-XL was seen. Apoptotic Index, telomerase activity and BRCA1 expression showed no correlation with any of the parameters. Our study confirms the fact that multiple gene interactions govern the pathogenesis of ovarian cancer, and analyzing ascitic cells of ovarian cancer patients may help to delineate molecular profile of the primary tumor.