<Emphasis Type="Italic">In vitro</Emphasis> study of HAX1 gene therapy by retro viral transduction as a therapeutic target in severe congenital neutropenia

Severe congenital neutropenia (SCN) is a primary immunodeficiency disease in which a number of underlying gene defects are responsible for abnormalities in neutrophil development. The HCLS1-associated protein X1 (HAX1) mutation is associated with an autosomal-recessive form of SCN. Considering the potential of gene therapy approaches for the treatment of monogenic disorders, in this study we aimed to develop retroviral vectors expressing coding sequences (CDS) to be used for the removal of the genetic blockade in deficient hematopoietic cells. Following amplification of CDS with primers containing appropriate restriction sites, HAX1 CDS was cloned into an intermediate vector using TA-cloning. The sequence was transferred into a retroviral vector, followed by retroviral packaging in Plat-A cells. To show HAX1 protein expression, HEK293T cells were exposed to 10 multiplicity of infection (MOI) of retroviral particles and HAX1 expression was confirmed in these cells, using indirect intracellular flow cytometry. This vector was applied for in vitro transduction of hematopoietic stem cell with HAX1 mutation; after 11 days, cultured cells were analyzed for CD66acde and CD177 (neutrophil surface markers) expression. Increased neutrophil production in HAX1 viral vector-expressing hematopoietic cells was observed as compared to control vector transduced cells. Hence, according to the results, this type of therapy could be considered a potential treatment protocol for the disease.     

Effect of lemon essential oil on the developmental stages of Trialeurodes vaporariorum West (Homoptera: Aleyrodidae)

Trialeurodes vaporariorum (Homoptera: Aleyrodidae), is a destructive pest of many ornamental and greenhouse crops throughout the world. In this research, effect of essential oil derived from lemon peel, Citrus aurantifolia (Hook.) on mortality of eggs, first-instar nymphs and on adult oviposition of T. vaporariorum (Westwood) was determined under laboratory conditions. Analysis of Citrus aurantifolia essential oil used for insect fumigation by phase gas chromatography revealed the presence of 16 compounds including Limonene (56.6%), β-pinene (16.3%) and α-terpineol (11.3%). Five concentrations of essential oil – 0.002, 0.004, 0.008, 0.016 and 0.032 (μl/ml) – were applied in fumigant toxicity experiments. Greater mortality was observed with increasing dose of essential oil. First-instar nymphs were more sensitive to essential oil treatments compared with eggs and adults that reduced the survival rate of T. vaporariorum by 58, 70 and 56% after treatment of eggs, nymphs and adult, respectively. Based on this study, essential oil derived from C. aurantifolia could be used as an effective and environmentally sustainable bioinsecticide for the control of T. vaporariorum.     

Bactericidal activity of nukacin ISK-1: an alternative mode of action

We previously reported bacteriostatic action of nukacin ISK-1 against Bacillus subtilis JCM 1465^T. Here, we found its bactericidal activity against Micrococcus luteus DSM 1790 and Staphylococcus simulans 22, showing decrease in cell viability, cell lysis, and dissipation of the membrane potential. Moreover, leakage of small molecules such as K⁺, suggested the formation of small-sized or specific K⁺-conducting-pores by nukacin ISK-1.     

Evolutionary history of the Persian Jird,Meriones persicus,based on genetics,species distribution modelling and morphometric data

The Persian Jird, Meriones persicus, is distributed from Eastern Anatolia to Afghanistan and western Pakistan. Six subspecies were described based on skull features and coat colours, but the validity of these subspecies is uncertain, and no molecular work has ever been conducted on this species. Iran appears to be a key geographical region in which to revise the systematic and evolutionary history of this species, because five of the six subspecies are present in this country. To evaluate the phylogeographical history and taxonomy of this species in Iran, we used a combination of genetic (cytochrome b gene sequences of 70 specimens) and geometric morphometric (2D landmarks on the ventral side of skull of 258 specimens) analyses. We also used ecological niche modelling to make inferences about the evolutionary history of these lineages. Our molecular data highlight the existence of four genetic lineages, but they only partly correspond to the previously described subspecies. Our molecular and morphometric data confirm the validity of M. p. rossicus and show that it has a wider geographical range than previously thought. M. p. gurganensis and M. p. baptistae are genetically very close. The skull of M. p. gurganensis is morphologically distinguishable from other subspecies. The subspecies M. p. persicus and M. p. baptistae are genetically distinct, but morphologically close. Meriones p. ambrosius is genetically close to M. p. persicus, and additional analyses with more specimens are needed to validate its subspecific status. The genetic structure observed in Iran seems to fit the topography and biogeography of the country and emphasize the role of the Abarkooh, Central and Lut deserts as barriers to gene flow. All intraspecific divergent events within the Persian Jird occurred during the last 1.4 My, suggesting that climatic changes probably trigger diversification within this species. Our genetic and species niche modelling results suggest that potential refugial areas persisted during glacial periods for this species in north‐western Zagros Mountains, north‐eastern Alborz Mountains and Kohrud Mountains.     

Inflammation,Aging, and Cancer: Tumoricidal Versus Tumorigenesis of Immunity

Acute inflammation is a highly regulated defense mechanism of immune system possessing two well-balanced and biologically opposing arms termed apoptosis (‘Yin’) and wound healing (‘Yang’) processes. Unresolved or chronic inflammation (oxidative stress) is perhaps the loss of balance between ‘Yin’ and ‘Yang’ that would induce co-expression of exaggerated or ‘mismatched’ apoptotic and wound healing factors in the microenvironment of tissues (‘immune meltdown’). Unresolved inflammation could initiate the genesis of many age-associated chronic illnesses such as autoimmune and neurodegenerative diseases or tumors/cancers. In this perspective ‘birds’ eye’ view of major interrelated co-morbidity risk factors that participate in biological shifts of growth-arresting (‘tumoricidal’) or growth-promoting (‘tumorigenic’) properties of immune cells and the genesis of chronic inflammatory diseases and cancer will be discussed. Persistent inflammation is perhaps a common denominator in the genesis of nearly all age-associated health problems or cancer. Future challenging opportunities for diagnosis, prevention, and/or therapy of chronic illnesses will require an integrated understanding and identification of developmental phases of inflammation-induced immune dysfunction and age-associated hormonal and physiological readjustments of organ systems. Designing suitable cohort studies to establish the oxido-redox status of adults may prove to be an effective strategy in assessing individual’s health toward developing personal medicine for healthy aging.     

Biliverdin Reductase Is Heat Resistant and Coexpressed with Constitutive and Heat Shock Forms of Heme Oxygenase in Brain

Abstract: Two heme oxygenase (HO) isozymes—HO-1, which is a heat shock protein (HSP32), and HO-2—catalyze the isomer-specific production of biliverdin IXα and carbon monoxide. The latter has the potential of functioning as a neurotransmitter, whereas the reduced form of biliverdin, bilirubin, has potent antioxidant activity. Formation of bilirubin is catalyzed by biliverdin reductase (BVR). The reductase is a unique enzyme in being dual pyridine nucleotide and dual pH dependent. Here, we show that the reductase is resistant to thermal stress at both the protein and message level. We further demonstrate that the reductase is coexpressed in cells that display HO-1 and/or HO-2 under normal conditions, as well as in regions and cell types that have the potential to express heat shock-inducible HO-1 protein. Exposure of male rats to 42°C for 20 min did not decrease brain BVR activity, but caused a slight increase in NADPH-and NADH-dependent activities at 1 and 6 h following hyperthermia. High levels of the ～ 1.5-kb BVR mRNA were detected in control brain; it too displayed thermal tolerance. Similarly, the pattern of multiplicity of net charge variants of the enzyme purified from brain of heat-shocked rats did not differ from the control pattern. Immunochemical localization of BVR protein in normal brain correlated well with the presence of HO-1 and/or HO-2 throughout the forebrain, diencephalon, cerebellum, and brainstem regions. There were select neuronal and nonneuronal cells in the substantia nigra and cerebellum that did express the reductase under normal conditions, wherein no HO isozymes could be detected. The same population, however, responded to heat shock by an intense increase in the level of HO-1. We postulate that the constitutive presence of the reductase in this cell population and the overall thermal stability of the enzyme represent a safeguard mechanism in the brain for the prompt conversion of biliverdin to bilirubin under conditions when oxidation of the heme moiety of denatured hemoproteins by HO-1 is accelerated.     

A fast model of voltage-dependent NMDA receptors

NMDA receptors are among the crucial elements of central nervous system models. Recent studies show that both conductance and kinetics of these receptors are changing voltage-dependently in some parts of the brain. Therefore, several models have been introduced to simulate their current. However, on the one hand, kinetic models—which are able to simulate these voltage-dependent phenomena—are computationally expensive for modeling of large neural networks. On the other hand, classic exponential models, which are computationally less expensive, are not able to simulate the voltage-dependency of these receptors, accurately. In this study, we have modified these classic models to endow them with the voltage-dependent conductance and time constants. Temperature sensitivity and desensitization of these receptors are also taken into account. We show that, it is possible to simulate the most important physiological aspects of NMDA receptor’s behavior using only three to four differential equations, which is significantly smaller than the previous kinetic models. Consequently, it seems that our model is both fast and physiologically plausible and therefore is a suitable candidate for the modeling of large neural networks.     

Likelihood of acute coronary syndrome in emergency department chest pain patients varies with time of presentation

ABSTRACT: BACKGROUND: There is a circadian and circaseptal (weekly) variation in the onset of acute coronary syndrome (ACS). The aim of this study was to elucidate whether the likelihood of ACS among emergency department (ED) chest pain patients varies with the time of presentation. METHODS: All patients presenting to the Lund ED at Skane University Hospital with chest pain or discomfort during 2006 and 2007 were retrospectively included. Age, sex, arrival time at the ED and discharge diagnose (ACS or not) were obtained from the electronic medical records. RESULTS: There was a clear but moderate circadian variation in the likelihood of ACS among presenting chest pain patients, the likelihood between 8 and 10 am being almost twice as high as between 6 and 8 pm. This was mainly explained by a variation in the ACS likelihood in females and patients under 65 years, with no significant variation in males and patients over 65 years. There was no significant circaseptal variation in the ACS likelihood. CONCLUSIONS: Our results indicate that there is a circadian variation in the likelihood of ACS among ED chest pain patients, and suggest that physicians should consider the time of presentation to the ED when determining the likelihood of ACS.     

Periocular Dirofilariasis in a Young Woman: A Case Report

A case of periocular dirofilariasis has been diagnosed in Iran. A 27-yr old female referred with a history of edema and redness in the left eye since 2 wk ago. On slit lamp examination, a moving thread-like worm was seen in the subconjunctival area. Two days later, a 126 mm long, live filaria worm was extracted from the lower lid subcutaneous tissue. The worm was diagnosed as, likelihood, Dirofilaria immitis on microscopic examinations. The present case suggests that D. immitis can cause periocular infections.