New insulins and newer insulin regimens: a review of their role in improving glycaemic control in patients with diabetes.

The legacy effect of early good glycaemic control in people with diabetes shows it is associated with reduction of microvascular and macrovascular complications. Insulin therapy is essential and lifesaving in individuals with type 1 diabetes and beneficial for those with type 2 diabetes who fail to achieve optimal glycaemic targets with other classes of glucose-lowering therapies. Since the introduction of insulin analogues, insulin management has changed. This follow-up review attempts to update our earlier publication from 2009 and discusses the role of new insulin analogues and newer insulin regimens. Recognising the advent of new quality and economic initiatives both in the UK and worldwide, this paper reviews current insulin prescribing and the pros and cons of prescribing analogues in comparison to the human insulins that are now gaining more acceptance in everyday clinical practice.

Phospholipid-binding proteins differ in their capacity to induce autoantibodies and murine systemic lupus erythematosus.

We have previously shown that immunization of nonautoimmune mice with the phospholipid-binding protein ß2-glycoprotein I (ß2GPI), in combination with lipopolysaccharide (LPS), induces a murine model of systemic lupus erythematosus (SLE), with sequential emergence of autoantibodies and glomerulonephritis. Here, we determine whether the paradigm for induction of murine SLE extends to other phospholipid-binding proteins. Mice were immunized with a phospholipid-binding protein (prothrombin (PT), protein S, or ß2GPI), or a nonphospholipid-binding protein (glu-plasminogen), in the presence of LPS. The breadth and degree of the autoantibody response, and the frequency of glomerulonephritis, varied among the three proteins, with ß2GPI being the most effective in inducing SLE-like disease. The phospholipid-binding proteins also differed in the pattern of serum cytokines they elicited. The most apparent difference between ß2GPI and the other phospholipid-binding proteins was in their ability to bind to LPS: ß2GPI bound to LPS, while PT and protein S did not. Our data suggest that binding to phospholipid(s) is a necessary, but not sufficient, condition for full induction of murine SLE. We propose that other properties, such as physiologic function, avidity for anionic phospholipids, and degree of interaction with other cell surface and/or circulating molecules (particularly LPS) may determine the range and severity of disease.

Expression of transglutaminase-2 isoforms in normal human tissues and cancer cell lines: dysregulation of alternative splicing in cancer.

The multiple enzymatic activities and functions of transglutaminase type 2 (TG2) may be attributed to alternative TG2 molecules produced by differential splicing of TG2 mRNA. Different RNA transcripts of the human TG2 gene (TGM2) have been identified, but the expression of TG2 multiple transcripts has never been systematically addressed. We have confirmed and rationalized the main TG2 variants and developed a screening assay for the detection of alternative splicing of TG2, based on real-time reverse-transcription PCR. We have quantified the multiple TG2 transcripts in a wide range of normal tissues and in cancer cell lines from four different sites of origin. Our data show a significant correlation in the expression of canonical and alternative TG2 isoforms in normal human tissue, but differences in alternative splicing of TG2 in cancer cell lines, suggesting that in cancer cells the alternative splicing of TG2 is a more active process.

Application of the paraboloidal model to assess mucosal changes following segmental intestinal transplantation in children.

Segmental living related small intestinal transplantation (LRSITx) is a therapeutic option for conditions that present with short gut syndromes. Recovery of small intestinal mucosa after transplantation is critical to function. We examined the posttransplant mucosal changes to understand the absorptive capabilities of transplanted small intestine. The study of human subjects is constrained by limited biopsy material; therefore, we developed a technique of villus area measurement by extrapolation from two-dimensional surgical biopsy images. Using a detailed model of the villus as the gold standard, two simpler models (cylindrical and paraboloid) were tested. Comparisons with the accurate measurement revealed that the cylinder model does not compare well in early posttransplant biopsies. The paraboloid function developed in this article worked very well under all conditions. The simplicity of the paraboloid model and its robustness made high-quality estimates of the absorptive surface area from abundant data relatively easy. The remodeling expected in the initial stages of growth was also captured by this model. Time-dependent villus growth curves were obtained for transplanted bowel. Serial biopsies showed an initial "reconstruction" dip around 2 weeks after transplant, followed by continuous growth of villus surface area. An eventual plateau resulted at an average of 6 months after transplant. This growth of villi was shown to parallel the improved absorption of electrolytes, amino acids, and water.

The role of recombinant factor VIIa for obstetric block in women with severe factor XI deficiency.

Regional blocks like spinal, epidural and combined spinal epidural (CSE) are relatively contraindicated in individuals with bleeding disorders. Consequently pregnant women with severe factor XI (FXI) deficiency are often denied this option during labour and caesarean section. We describe three women with severe FXI deficiency in whom regional block was performed with low-dose recombinant factor VIIa (rFVIIa) for their operative procedures during delivery. All women achieved haemostasis and had uncomplicated regional block, delivery and surgical procedures. The point of care device--rotational thromboelastometry (ROTEM) was used to monitor the patients' coagulation and determine the dose of rFVIIa to achieve in vitro haemostasis in these women that was then subsequently used in vivo. Low-dose rFVIIa seems to be effective and safe in the management of delivery and enables provision of regional blocks in women with severe FXI deficiency.

Nephroprotective and Nitric oxide Scavenging Activity of Tubers of Momordica tuberosa in Rats.

Hydroalcoholic extract (70% ethanol extract) of tubers of Momordica tuberosa Cogn. (Cucurbitaceae) was subjected to preliminary phytochemical screening by qualitative tests. Nitric oxide scavenging activity was performed by Griess reagent method. And nephroprotective activity was assessed in gentamicin, cisplatin and paracetamol induced renal damage in wistar rats (150-200 g) by standard methods. The protective property of 70% ethanol extract was assessed by measuring the levels of body weight, blood urea, serum creatinine, tissue glutathione and lipid peroxidation in administered doses. The extract exhibited free radical scavenging activity in dose dependant manner. And 100 µg/ml dose produced significantly higher scavenging activity than standard sodium metabisulphate at 25 µg/ml. Also, it significantly reduced the renal damage caused by cisplatin, gentamicin and paracetamol at a dose of 40 mg/kg.

Role of liquid membrane phenomenon in the anti-bacterial activity of Cefuroxime Sodium.

The role of liquid membrane phenomenon has been studied in the anti bacterial activity of cephalosporins i.e. Cefuroxime sodium. In our earlier publication [1] it was reported that hydraulic permeability data obtained to demonstrate the existence of liquid membrane in series with supporting membrane generated by Cefuroxime sodium. Transport of selected permeants (glucose, PABA, glycine, and ions like Mg(++), NH4 (+), PO4 (-), Ca(++), Na(+), K(+) and Cl(-)) through liquid membrane generated by Cefuroxime sodium in series with supporting membrane has been studied. The results indicated that the liquid membrane generated by Cefuroxime sodium inhibit the transport of various essential bio-molecules and permeants in to the cell. This modification in permeability of different permeants in the presence of the liquid membranes is likely to play significant role in the biological actions of Cefuroxime sodium. The anti-bacterial activity of Cefuroxime sodium further confirmed that the generation of liquid membrane by Cefuroxime sodium is also contributing for the antibacterial activity of them.

Antioxidant and hepatoprotective activity of tubers of Momordica tuberosa Cogn. against CCl4 induced liver injury in rats.

Hydro alcoholic extract of tubers of M. tuberosa was subjected to preliminary phytochemical screening and evaluated for in vitro and in vivo antioxidant and hepatoprotective activity against CCl4 induced liver damage in rats. Pretreatment with 70% ethanolic extract of M. tuberosa reversed CCl4 induced elevation of levels of serum biomarkers to near normal levels, suggesting that the tubers of M. tuberosa possess hepatoprotective property and this property may be attributed to the antioxidant property of the plant.

Hepatoprotective activity of Calotropis procera flowers against paracetamol-induced hepatic injury in rats.

Hydro-ethanolic extract (70%) of Calotropis procera flowers was prepared and tested for its hepatoprotective effect against paracetamol-induced hepatitis in rats. Alteration in the levels of biochemical markers of hepatic damage like SGPT, SGOT, ALP, bilirubin, cholesterol, HDL and tissue GSH were tested in both treated and untreated groups. Paracetamol (2 g/kg) has enhanced the SGPT, SGOT, ALP, bilirubin and cholesterol levels and reduced the serum levels of HDL and tissue level of GSH. Treatment with hydro-ethanolic extract of C. procera flowers (200 mg/kg and 400 mg/kg) has brought back the altered levels of biochemical markers to the near normal levels in the dose dependent manner.

Hepatoprotective and antioxidant activities of flowers of Calotropis procera (Ait) R. Br. in CCl4 induced hepatic damage.

Hepatoprotective activity of 70% ethanolic extract of flowers of C. procera was studied against CCl4 induced hepatic injury in albino rats and mice. In addition, antioxidant activity was studied by in vitro models. Pre-treatment with 70% ethanolic extract (CPA) reduced the biochemical markers of hepatic injury like serum glutamate pyruvate transaminase, serum glutamate oxaloacetate transaminase, alkaline phosphatase, bilirubin, cholesterol, HDL and tissue glutathione (GSH) levels. Similarly pretreatment with CPA reduced the CCl4 induced elevation in the pentobarbitone sleeping time. Histopathological observations also revealed that pretreatment with CPA protected the animals from CCl4 induced liver damage. CPA demonstrated dose dependant reduction in the in vitro and in vivo lipid peroxidation induced by CCl4. In addition it showed dose dependant free radical scavenging activity. The results indicate that flowers of C. procera possess hepatoprotective property possibly because of its anti-oxidant activity. This property may be attributed to the quercetin related flavonoids present in the flowers of Calotropis procera.

Practical aspects of planning, building, and interpreting tissue microarrays: the Cooperative Prostate Cancer Tissue Resource experience.

This is a review of several new approaches developed at or adopted by the Cooperative Prostate Cancer Tissue Resource (CPCTR) to resolve issues involved in tissue microarray (TMA) construction and use. CPCTR developed the first needle biopsy TMA, allowing researchers to obtain 200 or more consecutive cancer sections from a single biopsy core. Using radiographs of original paraffin blocks to measure tissue thickness we developed a method to produce TMAs with a larger number of usable sections. The modular approach to plan TMA construction is also a novel concept wherein TMAs of different types, such as tumor grade TMAs, metastasis TMA and hormone refractory tumors TMA can be combined to form an ensemble of TMAs with expanded research utility, such as support for tumor progression studies. We also implemented an open access TMA Data Exchange Specification that allows TMA data to be organized in a self-describing XML document annotated with well-defined common data elements. It ensures inter-laboratory reproducibility because it offers information describing the preparation of TMA blocks and slides. There are many important aspects that may be missed by both beginners and experienced investigators in areas of TMA experimental design, human subjects protection, population sample size, selection of tumor areas to sample, strategies for saving tissues, choice of antibodies for immunohistochemistry, and TMA data management.

Flow cytometry: a new approach for the molecular profiling of breast cancer.

The established method in prognosis of breast cancer includes detection of molecular markers, such as the estrogen receptor (ER), progesterone receptor (PR), and HER-2/neu. These markers are routinely checked via immunohistochemistry (IHC). HER-2/neu is also detected by fluorescent in situ hybridization (FISH). Flow cytometric analysis has not yet been used for detection of such markers. Flow cytometry was performed on four established breast cancer cell lines: MCF7, T47D, BT474, MDA-MB-231, and on one normal breast epithelial cell line: MCF10A. Flow cytometric analysis was used for the detection of ER, PR, HER-2/neu, epidermal growth factor receptor (EGFR), and E-cadherin. Currently, EGFR and E-cadherin are not standard predictive factors in determining survival of breast cancer patients, but both may be beneficial to prognosis. Cells undergoing flow cytometric analysis lost marker expression with increasing passage number. The highest expression was found at cells passaged 0-1 times. MCF7, T47D, and BT474 all had similar marker expression patterns. E-cadherin demonstrated a strongly positive pattern with marker expression of 85-92% among the three cell lines. ER, PR, and HER-2/neu demonstrated a weakly positive expression pattern when compared with E-cadherin. Marker expression ranged from 15 to 61%. These three cell lines were almost negative for expression of EGFR where expression ranged from 0 to 6%. MDA-MB-231 had almost no expression of all 5 markers, with positive values ranging from 0 to 5%. MCF10A had weak positive to almost negative expression values of ER, PR, HER-2/neu, and E-cadherin, which ranged from 3 to 13%. EGFR, both surface and cytoplasmic markers, again were not expressed in MCF10A cells with an expression value of <1%. We found that ER, PR, and HER-2/neu marker expressions in 5 out of 5 cell lines were consistent with established expression patterns. EGFR and E-cadherin expression in 4 out of 5 cell lines were also consistent with established expression patterns. We have shown that flow cytometry provides quantitative data on expression patterns of important prognostic markers in breast cancer.

Congenital esophageal stenosis: a case report and review of the literature.

A child with congenital esophageal stenosis (CES) demonstrates the need for accuracy in the diagnosis and management of this rare problem. Other etiologies of dysphagia must be looked into, but when the expected results are not realized CES should be considered. A range of diagnoses and diagnostic studies may help to get children with dysphagia the treatment that they need to resolve their symptoms.

The influence of implant design on accuracy of postoperative refraction.

PURPOSE: To assess the degree of accuracy of postoperative refraction that may be achieved with modern techniques and a new lens of modern design. METHODS: Five hundred eyes underwent refraction at one week and one month following phacoemulsification and implantation of the Centerflex lens (Rayner Intraocular Lenses Ltd style 570H). Results were compared with the target that had been computerised at the time of surgery, using the Hoffer Q formula for axial lengths less than 22.0 mm, the SRK-T for lengths greater than 24.5 mm and a mean of the two formulae for lengths between 22.0 and 24.5 mm. Statistical analysis used Fisher's exact and one way ANOVA tests. RESULTS: At one month 97% of refractions were within 1.0 D spherical equivalent (SE) of target and 81% within 0.5 SE. The mean absolute error was 0.37 +/- 0.39 SD. 75.8% of all eyes achieved 6/12 unaided and 88.6% achieved 6/9 with correction. In only 1.2% the refraction between one week and one month had altered by 0.5 D or more. CONCLUSIONS: The Centerflex when combined with modern methods of power calculation, can be associated with a high degree of accuracy in postoperative refraction.

Hypoglycemic activity of Abutilon indicum leaf extracts in rats.

Alcohol and water extracts of Abutilon indicum leaves (400 mg/kg, p.o.) showed significant hypoglycemic effect in normal rats 4 h after administration (23.10% and 26.95%, respectively).