Resveratrol may be beneficial in treatment of diabetic foot syndrome

Diabetic foot syndrome (DFS) is a late-stage complication of type 2 diabetes which originates from interplay among impaired tissue regeneration, vasculopathy, neuropathy and inflammation all on the background of insulin resistance. Despite astonishing mortality rate pharmacological approach in management of diabetic ulceration is almost non-existent. Foot pressure relief, wound debridement and infection control remain widely accepted options in the treatment of DFS. We hypothesize that resveratrol treatment and subsequent activation of SIRT1 pathway might be highly beneficial for patients with DFS. This prediction is based on multiple lines of evidence implicating resveratrol and sirtuins in restoration of insulin sensitivity, microcirculation, tissue regeneration, function of peripheral nerves and production of cytokines. Stabilized “nutraceutical” formulations of resveratrol with high absorption rate are essential to examine its potential medical benefits since dietary polyphenols are known to be rapidly metabolized by gut microflora and oxidized during absorption. Clinical trials with nutraceutical formulations and placebo are required to understand if resveratrol indeed holds the promise for treatment of DFS.     

Expression of class 3 semaphorins and their receptors in human breast neoplasia

Staton C A, Shaw L A, Valluru M, Hoh L, Koay I, Cross S S, Reed M W & Brown N J
(2011) Histopathology 59 , 274–282 Expression of class 3 semaphorins and their receptors in human breast neoplasia Aims: This study aimed to identify the involvement of class 3 semaphorins (Sema3) and receptors, neuropilins (Np1 and Np2) and plexins (A1–A4) in breast cancer development and angiogenesis. Methods and results: We quantified and correlated Sema3A, Sema3B, Sema3F and their known receptors and coreceptors Plexin‐A1, Plexin‐A3, Np1 and Np2 in sections of normal human breast, benign and pre‐malignant hyperplastic tissue, pre‐invasive and invasive cancer, and compared these findings with our previously published data on vascular endothelial growth factor (VEGF) and microvessel density (MVD) in the same samples. Histological analysis revealed that Sema3B was expressed more strongly and widely than Sema3A and 3F in normal breast tissue and all three semaphorins decreased with the transition from in situ to invasive cancer (P < 0.014). Plexin‐A3 decreased significantly with progression towards invasive cancer (P < 0.045), whereas Plexin‐A1 expression was only significantly reduced once invasion had occurred (P = 0.012). Np1 and Np2 were expressed in both endothelial and epithelial/tumour cells. Np2 expression did not change, but Np1 expression significantly increased in the spectrum from hyperplasia to ductal carcinoma in situ (P < 0.035), but decreased with invasive cancer. Conclusion: These data suggest that a decrease in class 3 semaphorin and their plexin receptors have some relationship with disease progression in ductal breast carcinoma.     

Postoperative intra-arterial methylene blue injection of colorectal cancer specimens increases the number of lymph nodes recovered

Törnroos A, Shabo I, Druvefors B, Arbman G & Olsson H
(2011) Histopathology 58 , 408–413
Postoperative intra‐arterial methylene blue injection of colorectal cancer specimens increases the number of lymph nodes recovered Aims: To determine the possible advantage of intra‐arterial injection of methylene blue with a view to improving lymph node recovery in postoperative examination of colorectal cancer specimens. Methods and results: Thirty‐two colorectal cancer specimens were assigned randomly to either dissection with intra‐arterial methylene blue injection or to routine dissection (without methylene blue injection). Immediately postoperatively, the specimens in the staining group were injected intra‐arterially with methylene blue dye. The two procedures were compared with respect to the number of lymph nodes recovered. The number of recovered lymph nodes was significantly higher in the intra‐arterial methylene blue injection group than in the group investigated with routine procedures (P < 0.0001). Conclusion: The intra‐arterial methylene blue injection method is fairly easy to use postoperatively and increases significantly the number of lymph nodes recovered in colorectal cancer specimens.     

Sequence-specific stalling of DNA polymerase γ and the effects of mutations causing progressive ophthalmoplegia

A large number of mutations in the gene encoding the catalytic subunit of mitochondrial DNA polymerase γ (POLγA) cause human disease. The Y955C mutation is common and leads to a dominant disease with progressive external ophthalmoplegia and other symptoms. The biochemical effect of the Y955C mutation has been extensively studied and it has been reported to lower enzyme processivity due to decreased capacity to utilize dNTPs. However, it is unclear why this biochemical defect leads to a dominant disease. Consistent with previous reports, we show here that the POLγA:Y955C enzyme only synthesizes short DNA products at dNTP concentrations that are sufficient for proper function of wild-type POLγA. In addition, we find that this phenotype is overcome by increasing the dNTP concentration, e.g. dATP. At low dATP concentrations, the POLγA:Y955C enzyme stalls at dATP insertion sites and instead enters a polymerase/exonuclease idling mode. The POLγA:Y955C enzyme will compete with wild-type POLγA for primer utilization, and this will result in a heterogeneous population of short and long DNA replication products. In addition, there is a possibility that POLγA:Y955C is recruited to nicks of mtDNA and there enters an idling mode preventing ligation. Our results provide a novel explanation for the dominant mtDNA replication phenotypes seen in patients harboring the Y955C mutation, including the existence of site-specific stalling. Our data may also explain why mutations that disturb dATP pools can be especially deleterious for mtDNA synthesis.     

NK cell function and antibodies mediating ADCC in HIV-1-infected viremic and controller patients

Natural killer (NK) cells have been suggested to play a protective role in HIV disease progression. One potent effector mechanism of NK cells is antibody-dependent cellular cytotoxicity (ADCC) mediated by antiviral antibodies binding to the FcγRIIIa receptor (CD16) on NK cells. We investigated NK cell-mediated ADCC function and the presence of ADCC antibodies in plasma from 20 HIV-1-infected patients and 10 healthy donors. The HIV-positive patients were divided into two groups: six who controlled viremia for at least 8 y without treatment (controllers), and 14 who were persistently viremic and not currently on treatment. Plasma from both patient groups induced NK cell IFN-γ expression and degranulation in response to HIV-1 envelope (Env) gp140-protein-coated cells. Patient antibodies mediating ADCC were largely directed towards the Env V3 loop, as identified by a gp140 protein lacking the V3 loop. Interestingly, in two controllers ADCC-mediating antibodies were more broadly directed to other parts of Env. A high viral load in patients correlated with decreased ADCC-mediated cytolysis of gp140-protein-coated target cells. NK cells from both infected patients and healthy donors degranulated efficiently in the presence of antibody-coated HIV-1-infected Jurkat cells. In conclusion, the character of ADCC-mediating antibodies differed in some controllers compared to viremic patients. NK cell ADCC activity is not compromised in HIV-infected patients.     

Biological significance of 5S rRNA import into human mitochondria: role of ribosomal protein MRP-L18

5S rRNA is an essential component of ribosomes of all living organisms, the only known exceptions being mitochondrial ribosomes of fungi, animals, and some protists. An intriguing situation distinguishes mammalian cells: Although the mitochondrial genome contains no 5S rRNA genes, abundant import of the nuclear DNA-encoded 5S rRNA into mitochondria was reported. Neither the detailed mechanism of this pathway nor its rationale was clarified to date. In this study, we describe an elegant molecular conveyor composed of a previously identified human 5S rRNA import factor, rhodanese, and mitochondrial ribosomal protein L18, thanks to which 5S rRNA molecules can be specifically withdrawn from the cytosolic pool and redirected to mitochondria, bypassing the classic nucleolar reimport pathway. Inside mitochondria, the cytosolic 5S rRNA is shown to be associated with mitochondrial ribosomes.     

Microfilariae in birds in the Czech Republic,including a note on adult nematodes <Emphasis Type="Italic">Eufilaria delicata</Emphasis> in a song thrush <Emphasis Type="Italic">Turdus philomelos</Emphasis>

Blood samples of more than 1,100 passerineform birds of 40 species were investigated for the occurrence of microfilariae. In the year 2005, 3 out of 677 birds of 31 species (prevalence 0.4%) were infected with microfilariae during the post-nesting period. During the pre-nesting period in the year 2007, 11 out of 438 birds of 31 species were infected with microfilariae (prevalence 2.5%). Both the pre-nesting and post-nesting examinations were conducted at the same location in the northeastern part of the Czech Republic. The microfilariae of the Eufilaria delicata and Ornithofilaria mavis species were found in Turdus merula, Turdus philomelos, and Erithacus rubecula (Passeriformes, Turdidae). Single individual of Poecile montanus (Passeriformes, Paridae) was infected with undetermined microfilariae. The morphometric variability of microfilariae found in T. philomelos, E. rubecula, and Poecile montanus were recorded. Infections caused by microfilariae E. delicata were more frequent than infections caused by O. mavis. Seven adult nematodes E. delicata were found in a subcutaneous cyst on the heel joint in one T. philomelos, which is the first record of adult E. delicata nematodes in birds in the Czech Republic.     

Immune response of chicken gut to natural colonization by gut microflora and to Salmonella enterica serovar enteritidis infection

In commercial poultry production, there is a lack of natural flora providers since chickens are hatched in the clean environment of a hatchery. Events occurring soon after hatching are therefore of particular importance, and that is why we were interested in the development of the gut microbial community, the immune response to natural microbial colonization, and the response to Salmonella enterica serovar Enteritidis infection as a function of chicken age. The complexity of chicken gut microbiota gradually increased from day 1 to day 19 of life and consisted of Proteobacteria and Firmicutes. For the first 3 days of life, chicken cecum was protected by increased expression of chicken β-defensins (i.e., gallinacins 1, 2, 4, and 6), expression of which dropped from day 4 of life. On the other hand, a transient increase in interleukin-8 (IL-8) and IL-17 expression could be observed in chicken cecum on day 4 of life, indicating physiological inflammation and maturation of the gut immune system. In agreement, the response of chickens infected with S. Enteritidis on days 1, 4, and 16 of life shifted from Th1 (characterized mainly by induction of gamma interferon [IFN-γ] and inducible nitric oxide synthase [iNOS]), observed in younger chickens, to Th17, observed in 16-day-old chickens (characterized mainly by IL-17 induction). Active modification of chicken gut microbiota in the future may accelerate or potentiate the maturation of the gut immune system and increase its resistance to infection with different pathogens.     

A closer look at RapidArc® radiosurgery plans using very small fields

RapidArc? has become the treatment of choice for an increasing number of treatment sites in many clinics. The extensive use of multiple subfields in RapidArc? treatments presents unique challenges, especially for small targets treated in few fractions. In this work, very small static fields and subsequently RapidArc? and conventional plans for two targets (0.4 and 9.9 cm(3)) were investigated. Doses from static fields 1-4 MLC leaves (0.25-1.00 cm) wide, and larger fields with 1-4 MLC leaves closed in their centres, were measured using the portal dosimeter-based QA system EPIQA (v?1.3) and gafchromic film. RapidArc and conventional plans for two tumours were then measured using EPIQA, gafchromic EBT2 film and the phantom-based QA system Delta4. Eclipse 8.6 and 8.9, grid spacings of 1.25 and 2.50 mm and a Varian HD linac were used. For static fields one MLC leaf wide, the dose was underestimated by Eclipse by as much as 53% (v?8.6, 2.5 mm grid). Eclipse underestimated the dose downstream from a few MLC leaves closed in the centre of a large MLC field by as much as 30%. Eclipse consistently overestimated the width of the penumbra by about 100%. For the conventional plans, there was good agreement between the calculated and measured dose for the 9.9 cm(3) PTV, but a 10% underdose was observed for the 0.4 cm(3) PTV. For the RapidArc? plans, the measured dose for the 9.9 cm(3) PTV was in good agreement with the calculated one. However, for the 0.4 cm(3) PTV about 10% overdosing was detected (Eclipse v 8.6, 2.5 mm grid spacing). EPIQA data indicated that the measured dose profiles were overmodulated compared to the calculated one. The use of small subfields, typically a few MLC leaves wide, or larger fields with one or a few MLC leaves closed in its centre can result in significant errors in the dose calculation. The detector systems used vary in their ability to detect the discrepancies. Using a smaller grid size and newer version of Eclipse reduces the discrepancies observed in this work but does not eliminate them.