Genome projects for the parasitic helminths Brugia malayi (a representative filarial nematode) and Schistosoma were initiated in 1995 by the World Health Organization with the ultimate objectives of identifying new vaccine candidates and drug targets and of developing low resolution genome maps. Because no genetic maps are available, and very few genes have been characterized from either parasite group, the first goal of both Initiatives has been to catalogue new genes for future placement on chromosome and physical maps. These genes have been identified by the expressed sequence tag (EST) approach, utilising cDNA libraries constructed from diverse life cycle stages. To date, the Initiatives have deposited over 16,000 Brugia ESTs and nearly 8000 Schistosoma ESTs in Genbank's dbEST database, corresponding to 6000 and over 3600 genes respectively (33% of Brugia's estimated gene compliment, 18-24% of that of Schistosoma). Large fragment, genomic libraries have been constructed in BAC and YAC vectors for studies of genomic organization and for physical and chromosome mapping, and public, hypertext genomic databases have been established to facilitate data access. We present a summary of progress within the helminth genome initiatives and give several examples of important gene discoveries and future applications of these data.

BACKGROUND

Pilonidal disease is common. Excessive hair growth in the natal cleft is thought to be a factor in initiating these sinuses. It is chronic and intermittent in nature and treatment can be difficult. Hair removal by shaving or use of creams is often advised as a compliment to surgical treatments. However, access to the natal cleft can be difficult. Laser removal of hair in the natal cleft is considered as an aid to healing the pilonidal sinus.

METHODS

Over a 5-year period, 14 patients with recurrent pilonidal disease were treated in our unit with laser depilation. They were all contacted by postal questionnaire, and those with ongoing disease were asked to return to the clinic for evaluation and possible further treatment.

RESULTS

All patients returned the postal questionnaire. Of the 14 patients, 4 had on-going disease and received further depilation with the Alexandrite laser. All are now healed with no reported complications. All patients found the procedure painful and received local anaesthetic.

CONCLUSIONS

Laser depilation in the natal cleft is by no means a cure for pilonidal disease. Removal of hair by this method represents an alternative and effective method of hair removal and, although long lasting, is only temporary. However, it allows the sinuses to heal rapidly. It is relatively safe, and simple to teach, with few complications. It should thus be considered as an aid to healing the problem pilonidal sinus.

BACKGROUND

Polymorphisms or mutations in hypoxia inducible factor-1 alpha (HIF-1alpha) that increases its activity and stability under normoxia have recently been identified. Likewise, disruption of the TSC1/TSC2 complex through loss of TSC1 or TSC2 has been shown to result in abnormal accumulation of HIF-1alpha. Here, we investigate the novel polymorphisms in exon 12, that approximate the oxygen-dependent degradation domain of HIF-1alpha in five cell lines and 28 patients with oral squamous carcinomas. Moreover, we assess for the presence of polymorphisms and mutations in TSC1 and TSC2, to ascertain if dysregulation of such might complement HIF-1alpha expression.

RESULTS

Denaturing high pressure liquid chromatography (DHPLC) analysis on PCR fragments in exon 12 of HIF-1alpha from 28 patients with OSCC revealed that 6 of 28 patients had mismatched heteroduplex patterns. Genomic DNA was extracted from peripheral blood leukocytes and direct sequencing showed that in 5 of the six cases these changes represented polymorphisms while, one case was a somatic mutation. Analyses of TSC1 and TSC2 revealed heteroduplexes in exons: TSC1 exon 17; TSC2 exons 36, 40, and 41. The relative levels of HIF-1alpha were significantly greater for tumors possessing a HIF-1alpha polymorphism or mutation within exon 12, whereas tumors possessing a deletion or polymorphism in TSC1/TSC2 displayed a trend for higher levels of HIF-1alpha. Western blot analyses for HIF-1alpha, TSC1 and TSC2 in five SCC cell lines revealed high levels of HIF-1alpha in SCC cells possessing TSC1 and/or TSC2 mutations. Wild-type TSC2 cells targeted with siRNA to TSC2 exhibited increased levels of HIF-1alpha. Transfection of a HIF-1alpha mutant produced higher levels of HIF-1alpha in TSC1/TSC2 mutant cell lines than in wild type cells. TSC1/TSC2 mutant cell lines administered Rapamycin blocked S6 phorphorylation and diminished the levels of HIF-1alpha to those observed in cell lines with wild type TSC1/TSC2.

CONCLUSIONS

Dysregulation of the TSC1/TSC2 complex by mutation compliments HIF-1alpha polymorphisms in the expression of HIF-1alpha in SCC of the head and neck, and may provide biomarkers to predict responses to specific therapies and overall disease prognosis.

BACKGROUND

There is a need for a standardized, impartial calibration, and validation protocol on confocal spectral imaging (CSI) microscope systems. To achieve this goal, it is necessary to have testing tools to provide a reproducible way to evaluate instrument performance.

METHODS

We evaluated fluorescent spectral beads (FocalCheck) from Molecular Probes/Invitrogen that consist of four pairs with emissions between 500 and 725 nm and a europium macrocycle quantum dye bead. These bead tools compliment our previously published protocol for testing spectral imaging systems that used an inexpensive multi-ion discharge lamp (MIDL) that contains Hg(+), Ar(+), and inorganic fluorophores that emits distinct, stable spectral features.

RESULTS

We acquired the spectra of the FocalCheck beads on a Zeiss 510 Meta, a Leica TCS-SP1, a Leica SP2 AOBS, an Olympus FV 1000, and a Nikon C1Si confocal systems and a PARISS microscopic spectral system and of the europium beads on the Leica TCS-SP1 and PARISS spectral imaging systems. A lack of performance with some equipment between 650 and 750 nm was identified using the far red pair of the FocalCheck beads. The position of the slider in front of PMT 2 that reflects light into PMT 1 and PMT 3 affected the measurement of all bead intensities. Unmixing algorithms were used to separate beads with different fluorochromes and separate two fluorochromes on the same bead.

CONCLUSIONS

The FocalCheck multi-spectral beads yielded similar profiles on four CSI systems and a PARISS spectral system. The utilization of the spectral FocalCheck beads is helpful to evaluate proper spectral performance, especially in the far red region. Europium beads provide a very narrow spectrum that can help to identify machines that have spectral problems. The dyes located on individual beads or mixed together in ring-core configuration can be used as test particles to demonstrate spectral unmixing with various algorithms.

BACKGROUND

Patient-reported outcomes (PROs) are of growing importance in research and clinical care and may be used as primary outcomes or as compliments to traditional surgical outcomes. In assessing the impact of surgical and traumatic scars, PROs are often the most meaningful. To assess outcomes from the patient perspective, rigorously developed and validated PRO instruments are essential.

METHODS

The authors conducted a systematic literature review to identify PRO instruments developed and/or validated for patients with surgical and/or non-burn traumatic scars. Identified instruments were assessed for content, development process, and validation under recommended guidelines for PRO instrument development.

RESULTS

The systematic review identified 6534 articles. After review, we identified four PRO instruments meeting inclusion criteria: patient and observer scar assessment scale (POSAS), bock quality of life questionnaire for patients with keloid and hypertrophic scarring (Bock), patient scar assessment questionnaire (PSAQ), and patient-reported impact of scars measure (PRISM). Common concepts measured were symptoms and psychosocial well-being. Only PSAQ had a dedicated appearance domain. Qualitative data were used to inform content for the PSAQ and PRISM, and a modern psychometric approach (Rasch Measurement Theory) was used to develop PRISM and to test POSAS. Overall, PRISM demonstrated the most rigorous design and validation process, however, was limited by the lack of a dedicated appearance domain.

CONCLUSIONS

PRO instruments to evaluate outcomes in scars exist but vary in terms of concepts measured and psychometric soundness. This review discusses the strengths and weaknesses of existing instruments, highlighting the need for future scar-focused PRO instrument development.

METHODS

This journal requires that authors assign a level of evidence to each article. For a full description of these Evidence-Based Medicine ratings, please refer to Table of Contents or the online Instructions to Authors www.springer.com/00266 .

The American Association of Physicists in Medicine Task Group 36 (AAPM TG-36) data can be used to estimate peripheral dose (PD) distributions outside the primary radiation field. However, the report data does not apply to linear accelerators equipped with a multileaf collimator (MLC) and universal wedge (UW). Tertiary multileaf collimators have been shown to significantly affect PD distributions and TG-36 reported data. Measurements were performed to evaluate PD distributions for a linear accelerator equipped with a secondary MLC, backup diaphragms, and UW. This data can be used to compliment the TG-36 report for estimation of doses to critical structures outside primary radiation fields. For the evaluated linear accelerator, an MLC is incorporated in the upper secondary collimator jaws. Backup shielding diaphragms are located underneath the MLC. At the nominal collimator position, the MLC and the backup diaphragm provide collimation primarily in the transverse direction. Conventional, solid tungsten-alloy jaws, located underneath the backup diaphragms, provide secondary collimation in the longitudinal direction. The universal wedge provides dose modulation in the direction of the conventional jaws. Measurements were made with an ionization chamber inserted into a 20x40x120 cm(3) water-equivalent plastic phantom with the secondary collimator and MLC settings of 5x5, 10x10, 15x15, and 25x25 cm(2) with and without UW. Data was acquired along the machine's longitudinal axis for 6, 10, and 18 MV photons. Peripheral dose distributions were measured with the collimator rotated to 0 degrees and 270 degrees for open field measurements and to 0 degrees, 180 degrees, and 270 degrees for wedged fields (IEC 1217). This allowed evaluation of peripheral dose distributions as a function of collimator rotation. Wedged fields were normalized to deliver the same dose at the depth of maximum dose on the central axis as open fields. The measured PD distributions were generally comparable to data reported by TG-36. At distances close to the field edge (less than 30 or 40 cm), the measured PD distributions were lower when the measurement point was shielded by solid jaws than with MLC and backup diaphragm. At longer distances, this trend reversed for all energies and evaluated field sizes. However, the difference in PD distribution with collimator rotation was not large enough to warrant strategic positioning of the collimator to reduce dose to critical structures outside the primary radiation field. Because internal scatter dominates close to the field edge, wedged PD distributions were comparable to open field doses at distances closer than 30 cm. However, at distances larger than 30 cm from the field edge, wedged PD distributions were significantly grater than those for open fields due to increased contribution of leakage radiation. Increased leakage radiation is due to the increase in wedged field monitor units, which is related to a small wedge factor (0.27 to 0.29).

METHODS

Patients with positive smears for acid-fast bacilli were enrolled at tuberculosis (TB) clinics in the Khartoum region of Sudan.

OBJECTIVE

To identify the presence of drug resistant genotypes in M. tuberculosis isolates which are difficult to treat.

METHODS

Genus specific PCR-SSCP was performed to confirm the presence of M. tuberculosis in clinical isolates. Genotypic drug resistance testing was performed by mutation analysis and spoligotyping was used to monitor transmission and to identify epidemic strains.

RESULTS

Fifty (48%) of the original 105 samples were classified as M. tuberculosis. Four (4%) of the samples were typed as mycobacteria other than TB, while the remaining (n =50) samples were refractory to further molecular analysis. The fifty amplifiable M. tuberculosis samples were used for subsequent mutation analysis and typing. Mutations were identified in the genes conferring resistance to INH (kat G, 12%), RIF (rpoB, 8%), SM (r psL and rrs, 30%) and EMB (embB, 4%). Two of the samples (4%) had mutations in genes associated to both INH and RIF and can be classified as MDR-TB. Thirty-three percent (13/39) of the persistant tuberculosis cases (5/18 treatment failure; 5/14 relapse; 3/7 defaulter) had mutations accounting for drug resistance. A total of 27 different spoligotypes were identified from 49/50 samples. Twenty-nine (59%) of the isolates were grouped into one of seven clusters, while 20 (41%) showed unique patterns. One patient was infected with M. bovis.

CONCLUSIONS

This is the first molecular approach to characterize clinical isolates of M. tuberculosis from Sudan. The results show that drug resistance is indeed a serious problem and it may compliment the efforts of the National Tuberculosis Programme to improve strategies to control this disease.

The importance of natural enemies as the foundation of integrated pest management (IPM) is widely accepted, but few studies conduct the manipulative field experiments necessary to directly quantify their impact on pest populations in this context. This is particularly true for predators. Studying arthropod predator-prey interactions is inherently difficult: prey items are often completely consumed, individual predator-prey interactions are ephemeral (rendering their detection difficult) and the typically fluid or soft-bodied meals cannot be easily identified visually within predator guts. Serological techniques have long been used in arthropod predator gut-contents analysis, and current enzyme linked immunosorbent assays (ELISA) are highly specific and sensitive. Recently, polymerase chain reaction (PCR) methods for gut-contents analysis have developed rapidly and they now dominate the diagnostic methods used for gut-contents analysis in field-based research. This work has identified trophic linkages within food webs, determined predator diet breadth and preference, demonstrated the importance of cannibalism and intraguild predation within and between certain taxa, and confirmed the benefits (predator persistence) and potential disadvantages (reduced feeding on pest species) of the availability of alternative nonpest prey. Despite considerable efforts to calibrate gut-contents assays, these methods remain qualitative. Available techniques for predator gut-contents analysis can provide rapid, accurate, cost-effective identification of predation events. As such, they perfectly compliment the ecological methods developed to directly assess predator impacts on prey populations but which are imperfect at identifying the key predators. These diagnostic methods for gut-contents analysis are underexploited in agricultural research and they are almost never applied in unison with the critical field experiments to measure predator impact. This paper stresses the need for a combined approach and suggests a framework that would make this possible, so that appropriate natural enemies can be targeted in conservation biological control.

Rheumatoid arthritis is a progressive autoimmune disease characterised by severely swollen and painful joints. To compliment pharmacotherapy, people living with rheumatoid arthritis often turn to dietary interventions such as the Mediterranean diet. The aim of the present systematic review is to discuss the effects of the Mediterranean diet on the management and prevention of rheumatoid arthritis in human prospective studies. Four studies met the inclusion criteria, including two intervention studies reporting improvement in the pain visual analogue scale (p < 0.05) and a decrease in the health assessment questionnaire for rheumatoid arthritis score (p < 0.05) in the Mediterranean diet groups. Only one study reported a reduction in the 28 joint count disease activity score for rheumatoid arthritis for the Mediterranean diet group (p < 0.05). This review has identified beneficial effects of the Mediterranean diet in reducing pain and increasing physical function in people living with rheumatoid arthritis. However, there is currently insufficient evidence to support widespread recommendation of the Mediterranean diet for prevention of rheumatoid arthritis.

Zebrafish are a relevant and useful vertebrate model species to study normal- and patho-physiology, including that of the heart, due to conservation of protein-coding genes, organ system organisation and function, and efficient breeding and housing. Their amenability to genetic modification, particularly compared to other vertebrate species, is another great advantage, and is the focus of this review. A vast number of genetically engineered zebrafish lines and methods for their creation exist, but their incorporation into research programs is hindered by the overwhelming amount of technical details. The purpose of this paper is to provide a simplified guide to the fundamental information required by the uninitiated researcher for the thorough understanding, critical evaluation, and effective implementation of genetic approaches in the zebrafish. First, an overview of existing zebrafish lines generated through large scale chemical mutagenesis, retroviral insertional mutagenesis, and gene and enhancer trap screens is presented. Second, descriptions of commonly-used genetic modification methods are provided including Tol2 transposon, TALENs (transcription activator-like effector nucleases), and CRISPR/Cas9 (clustered regularly interspaced short palindromic repeats/CRISPR-associated protein 9). Lastly, design features of genetic modification strategies such as promoters, fluorescent reporters, and conditional transgenesis, are summarised. As a comprehensive resource containing both background information and technical notes of how to obtain or generate zebrafish, this review compliments existing resources to facilitate the use of genetically-modified zebrafish by researchers who are new to the field.

BACKGROUND

Engagement in regular exercise routinely is recommended as an intervention for managing and preventing migraine, and yet empirical support is far from definitive. We possess at best a weak understanding of how aerobic exercise and resulting change in aerobic capacity influence migraine, let alone the optimal parameters for exercise regimens as migraine therapy (eg, who will benefit, when to prescribe, optimal types, and doses/intensities of exercise, level of anticipated benefit). These fundamental knowledge gaps critically limit our capacity to deploy exercise as an intervention for migraine.

BACKGROUND

Clear articulation of the markers and mechanisms through which aerobic exercise confers benefits for migraine would prove invaluable and could yield insights on migraine pathophysiology. Neurovascular and neuroinflammatory pathways, including an effect on obesity or adiposity, are obvious candidates for study given their role both in migraine as well as the changes known to accrue with regular exercise. In addition to these biological pathways, improvements in aerobic fitness and migraine alike also are mediated by changes in psychological and sociocognitive factors. Indeed a number of specific mechanisms and pathways likely are operational in the relationship between exercise and migraine improvement, and it remains to be established whether these pathways operate in parallel or synergistically. As heuristics that might conceptually benefit our research programs here forward, we: (1) provide an extensive listing of potential mechanisms and markers that could account for the effects of aerobic exercise on migraine and are worthy of empirical exploration and (2) present two exemplar conceptual models depicting pathways through which exercise may serve to reduce the burden of migraine.

CONCLUSIONS

Should the promise of aerobic exercise as a feasible and effective migraine therapy be realized, this line of endeavor stands to benefit migraineurs (including the many who presently remain suboptimally treated) by providing a new therapeutic avenue as an alternative or augmentative compliment to established interventions for migraine.

Inhibin B is a glycoprotein produced predominantly by Sertoli cells which regulates pituitary FSH release by a negative feedback loop. The regulation of inhibin B is complex with changes in the pattern of secretion occurring during development, and many factors such as FSH, testosterone, Sertoli cell proliferation and germ cell complement likely to contribute to overall production. Systemic inhibin B concentrations seem to reflect the extreme ends of spermatogenic status with high levels of inhibin B observed in normal, fertile individuals and lower levels of inhibin B in individuals with severe damage to the testis as a result of germ cell depletion. Inhibin B has proved valuable in epidemiological studies exploring male infertility with data showing that inhibin B combined with FSH measurements has a higher positive predictive value for detecting male infertility than either alone. Inhibin B is proposed as a potential biomarker of testicular toxicity in rodent toxicity studies to compliment traditional endpoints. In pharmaceutical development, inhibin B might allow better linkage between animal study results and subsequent monitoring of testicular function in clinical trials. An international, intercompany project has been initiated to evaluate the overall suitability and limitations of inhibin B as a biomarker of testicular toxicity.

A number of intermittent team sports require that two consecutive periods of play (lasting for ~30-45 min) are separated by a 10-20 min half-time break. The half-time practices employed by team-sports players generally include returning to the changing rooms, temporarily relaxing from the cognitive and physical demands of the first half, rehydration and re-fuelling strategies, addressing injury or equipment concerns, and receiving tactical instruction and coach feedback. However, the typically passive nature of these actions has been associated with physiological changes that impair performance during the second half. Both physical and cognitive performances have been found to decline in the initial stages of subsequent exercise that follows half-time. An increased risk of injury has also been observed during this period. Therefore, half-time provides sports scientists and strength and conditioning coaches with an opportunity to optimise second-half performance. An overview of strategies thought to benefit team-sports athletes is presented; specifically, the efficacy of heat maintenance strategies (including passive and active methods), post-activation potentiation, hormonal priming, and modified hydro-nutritional practices are discussed. A theoretical model of applying these strategies in a manner that compliments current practice is also offered.

A need to reach poor blacks and Hispanics in Dade County, Florida, with information about the prevention and early detection of cancer was documented in data from the Florida Cancer Data System for 1981 through 1983. This information was brought to the target population through community media, churches, and agencies serving the poor. Guidelines for productive presentations evolved during the 4-year experience. A significant portion of the community was recruited for mammograms. A significant change to in situ and local stage of disease occurred among the screened group as compared to an unscreened cohort treated at the same facility. An estimated savings of about $9700 in treatment costs per cancer found has shown that mammography of the poor community saves money as well as lives. The establishment of a One-Stop Breast Diagnostic Center compliments the screening program and removes many barriers in reaching the goal of early cancer detection among the poor.

OBJECTIVE

To determine the load transfer patterns of femurs in the intact, immediate post-operative and long-term (remodelled) post-operative implanted conditions for Lubinus SPII and Müller-Curved cemented hip prostheses, and to examine to what extent remodelling may influence the long-term outcome.

METHODS

Experimental and finite element (FE) methods were applied to composite femurs under loads representing the heel-strike phase of gait, determining cortical bone and cement strains for the different femur conditions.

BACKGROUND

The authors previously developed protocols to measure bone and cement strains, and to produce remodelled femur specimens, yet these have not been applied together to compare strain patterns of different femur conditions. The Lubinus SPII is clinically more successful than the Müller-Curved stem, with failure mainly due to aseptic loosening.

METHODS

Cortical bone strains were determined in intact femurs. Six femurs each were implanted with the two stem types and cortical bone and cement strains were measured. Bone remodelling was recreated using a validated CAD-CAM procedure to remove a layer of proximal cortical bone, replicating a typical scenario found in stable clinical retrievals. Strains were remeasured. FE methods were used to compliment the experiments.

RESULTS

Stress shielding was reduced with remodelling, though bone strains did not return to their intact values, particularly around the calcar. Cement strains increased with remodelling. Differences occurred between the two stems; the Müller-Curved produced a more severe strain transition.

CONCLUSIONS

Procedures were successfully combined together to investigate in vitro the performance of two cemented stems, in immediate and long-term post-operative conditions. The increase of cement strains with remodelling is a potential indicator for in vivo cement failure.

CONCLUSIONS

The consequences of femoral bone remodelling on the long-term success of joint replacements are not well understood, where remodelling may lead to increased bone and cement stresses.

The absolute CD4+ T cell count in adults and CD4+ T cell percentage of lymphocytes (CD4%) in pediatrics compliment clinical history and physical examination to inform decisions about initiating antiretroviral therapy (ART). In addition, these immunologic markers predict host susceptibility to specific opportunistic infections, selected drug toxicities, and mortality. These benefits argue strongly for the availability of CD4+ T cell testing capacity in all settings where HIV infection is treated. Several currently available flow cytometry-based devices, and novel CD4+ T cell enumeration techniques such as the panleucogating CD4 are especially suitable for resource-constrained settings. At this time, unfortunately, the landscape of HIV care in sub-Saharan Africa is a mosaic characterized by large areas where CD4+ T cell testing capacity is limited or unavailable, and small, but growing, pockets where the capacity exists. Routine HIV quantification is currently unaffordable and unsustainable in the great majority of the region; therefore, a reliance on CD4+ T cell testing is inevitable for now. To this end, correcting the disparities in CD4+ T cell testing capacity and defining the minimum laboratory requirements for the safe use of antiretroviral drugs through well-designed clinical studies are some of the most urgent priorities of the ongoing global scale-up of ART.

Transrectal ultrasound was performed in 20 patients with suspected local pelvic recurrence after a radical pelvic operation: 9 had undergone radical prostatectomy and 11 had undergone radical cystoprostatectomy. Transrectal sonography verified the presence of recurrence in 19 of 20 patients (95%) and this was confirmed by biopsy of the visualized lesions. Analysis of the sonographic echo patterns encountered revealed that in 14 of 19 recurrent neoplasms (71.5%) the echogenic pattern was hypoechoic. In the remaining 6 patients (31.5%) the echo pattern was isoechoic. No hyperechoic lesions were noted. Based upon our findings and because of the low costs compared to other diagnostic modalities transrectal ultrasound represents an ideal technique to compliment the digital rectal examination in evaluation of patients suspected of harboring a local pelvic recurrence after a radical pelvic operation.

OBJECTIVE

This study aimed to examine differences in brain activation for various types of reward and feedback in adolescent Internet addicts (AIA) and normal adolescents (NA) using functional magnetic resonance imaging (fMRI).

METHODS

AIA (n = 15) and NA (n = 15) underwent fMRI while performing easy tasks for which performance feedback (PF), social reward (SR) (such as compliments), or monetary reward (MR) was given. Using the no reward (NR) condition, three types of contrasts (PF-NR, SR-NR, and MR-NR) were analyzed.

RESULTS

In NA, we observed activation in the reward-related subcortical system, self-related brain region, and other brain areas for the three contrasts, but these brain areas showed almost no activation in AIA. Instead, AIA showed significant activation in the dorsolateral prefrontal cortex for the PF-NR contrast and the negative correlation was found between the level of activation in the left superior temporal gyrus (BA 22) and the duration of Internet game use per day in AIA.

CONCLUSIONS

These findings suggest that AIA show reduced levels of self-related brain activation and decreased reward sensitivity irrespective of the type of reward and feedback. AIA may be only sensitive to error monitoring regardless of positive feelings, such as sense of satisfaction or achievement.

Although people with low self-esteem (LSEs) doubt their value to their romantic partners, they tend to resist positive feedback from their partners. This resistance undermines their relationships and has been difficult to overcome in past research. The authors investigated whether LSEs could be induced to take their partners' kind words to heart by manipulating how abstractly they described a recent compliment. In 3 studies, LSEs felt more positively about the compliments, about themselves, and about their relationships--as positively as people with high self-esteem (HSEs) felt--when they were encouraged to describe the meaning and significance of the compliments. The effects of this abstract meaning manipulation were still evident 2 weeks later. Thus, when prompted, LSEs can reframe affirmations from their partners to be as meaningful as HSEs generally believe them to be and, consequently, can feel just as secure and satisfied with their romantic relationships.

The 2008 World Health Organization (WHO) proposed revision of the classification of MDS recognizes a deletion (5q) subtype with mutation of Janus kinase-2 (JAK2(V617F)). We investigated the clonal origin of this gene mutation in a patient with del(5q) MDS presenting with thrombocytosis and normal hemoglobin. Analysis of colony forming units-granulocyte-monocyte (CFU-GM) and erythropoietin-independent growth of bone marrow (BM) and peripheral blood (PB) burst forming units-erythroid (BFU-E) showed that del(5q) and JAK2(V617F) existed in progenitors derived from independent clones. Fifty percent of endogenous erythroid colonies (EEC) harbored the JAK2(V617F) mutation whereas fluorescent in situ hybridization (Fish) with a chromosome 5 (q31.1) probe showed only a diploid allele compliment. Assessment of transcriptional clonality by iduronate-2-sulfatase (IDS) gene polymorphism suggested that JAK2(V617F) was acquired in at least two independent multipotent stem cell progeny. Our findings indicate that JAK2(V617F) mutant clones may arise in genetically discordant clones independent of del(5q).

Fibro/Adipogenic Progenitors (FAPs) are a multipotent progenitor population resident in skeletal muscle. During development and regeneration, FAPs provide trophic support to myogenic progenitors that is required for muscle fiber maturation and specification. FAPs also represent a major cellular source of fibrosis in degenerative disease states, highlighting them as a potential cellular target for anti-fibrotic muscle therapies. Effective and reproducible methods to isolate and culture highly purified FAP populations are therefore critical to further understand their biology. Here, we describe a fluorescent activated cell sorting (FACS) based protocol to isolate CD31-/CD45-/Integrin-α7-/Sca1+ FAPs from murine skeletal muscle including details of tissue collection and enzymatic muscle digestion. We also incorporate optimized methods of expanding and differentiated FAPs in vitro. Together, this protocol provides a complete workflow to study skeletal muscle derived FAPs and compliments downstream analytical, drug screening, and disease modeling applications.

Maternal mortality is one of the important indicators used for the measurement of maternal health. Although maternal mortality ratio remains high, maternal deaths in absolute numbers are rare in a community. To overcome this challenge, maternal near miss has been suggested as a compliment to maternal death. It is defined as pregnant or recently delivered woman who survived a complication during pregnancy, childbirth or 42 days after termination of pregnancy. So far various nomenclature and criteria have been used to identify maternal near-miss cases and there is lack of uniform criteria for identification of near miss. The World Health Organization recently published criteria based on markers of management and organ dysfunction, which would enable systematic data collection on near miss and development of summary estimates. The prevalence of near miss is higher in developing countries and causes are similar to those of maternal mortality namely hemorrhage, hypertensive disorders, sepsis and obstructed labor. Reviewing near miss cases provide significant information about the three delays in health seeking so that appropriate action is taken. It is useful in identifying health system failures and assessment of quality of maternal health-care. Certain maternal near miss indicators have been suggested to evaluate the quality of care. The near miss approach will be an important tool in evaluation and assessment of the newer strategies for improving maternal health.

Diabetes mellitus, the major cardiovascular risk factor, accentuates the inflammation and neovascularization processes leading to enhanced progression of atherosclerotic complications. Inflammation in diabetes mellitus is the key initiator of atherosclerotic process, which results in acute coronary events. Atherosclerosis evolves from the endothelial cell dysfunction and succeeding entry of hemodynamically derived leukocytes by migration, activation and production of lipid gruel leading to atheromatous plaque progression and subsequent regression. Diabetic plaque progression is associated with increased neovascularization, which is a nature's compliment in the sustenance of plaque growth by its nutrient supply. Neovessels may act as conduit for lipid debridment and alternative channel for inflammatory process. In addition, neovascularization induces intra-plaque hemorrhage due to the fragility of the neovessels and associated inflammation, resulting in plaque instability. The intra-plaque hemorrhage is a detrimental base, which begets the progress of atheroma by inducing oxidative stress and endothelial dysfunction. Intra-plaque hemorrhage is increased in diabetes with an associated increase in hemoglobin-haptoglobin complex (Hb-Hp2-2), which further induces oxidative stress and endothelial cell dysfunction. We conclude that inflammation and neovascularization of the plaque may act as major mechanism augmenting plaque instability in diabetes mellitus.