Stroke victims tend to prioritize speaking, writing, and walking as the three most important rehabilitation goals. Of note is that two of these goals involve communication. This underscores the significance of developing successful approaches to aphasia treatment for the several hundred thousand new aphasia patients each year and over 1 million stroke survivors with chronic aphasia in the U.S. alone. After several years of growth as a research tool, non-invasive brain stimulation (NBS) is gradually entering the arena of clinical aphasiology. In this review, we first examine the current state of knowledge of post-stroke language recovery including the contributions from the dominant and non-dominant hemispheres. Next, we briefly discuss the methods and the physiologic basis of the use of inhibitory and excitatory repetitive transcranial magnetic stimulation (rTMS) and transcranial direct current stimulation (tDCS) as research tools in patients who experience post-stroke aphasia. Finally, we provide a critical review of the most influential evidence behind the potential use of these two brain stimulation methods as clinical rehabilitative tools. Aphasia, defined as an impaired ability to communicate, is one of the most feared symptoms of stroke. About 21–38% of acute stroke survivors suffer from aphasia , new strategies e.g., adjuvant therapies, are required to boost recovery, especially in the chronic stages of stroke. While SLT is the most commonly employed treatment of aphasia, its therapeutic effects are quite variable and are generally modest  assessed language recovery mechanisms in a group of chronic aphasic patients by comparing blood-oxygen-level-dependent (BOLD) signal changes over a period of 10 weeks  derive from an understanding of the mechanisms of healthy brain function and neurological dysfunction after a brain injury, and (2) improve not only behavioral or cognitive performance but also the function of neural systems, which translates into favorable outcomes on everyday quality of life . Typically, low-frequency rTMS (<5 Hz) is characterized by decreased cortical excitability, whereas high-frequency rTMS (≥5 Hz) is characterized by enhanced excitability  for 20–40 min a day over 10–15 days, on sites in the right hemisphere that were homotopic to left hemisphere sites in the fronto-temporal language network , Aachen Aphasia Test (AAT), or Western Aphasia Battery (WAB)], Boston Naming Test (BNT), and accuracy and RT in picture naming tasks , a site in the IFG, was most frequently stimulated, some studies adopted a site-finding protocol Figure  either aApplication of optimal site-finding protocols, either rTMS- or neuroimaging-driven, is likely an improvement over blinded application of therapeutic rTMS as it accounts for individual variability in clinical factors such as lesion size and volume that could differentially influence the mechanisms of neuroplasticity in each patient  saline-soaked surface electrodes  to the scalp by a battery-driven device connected to two large ] and language batteries including picture naming tasks such as the BNT. Monti et al.  were firNaming accuracy also improved in 10 chronic aphasic patients, who received 1 mA a-tDCS on intact left frontal areas for 20 min per day for 5 days  and use ecologically valid outcome measure, several parameters in study design and stimulation methodology could be improved. First, except one study  and intensity of stimulation (>1 mA), c-tDCS may behave more like a-tDCS, in that it increases, rather than decreasing, the cortical excitability  after therapeutic rTMS as no such studies have yet been carried out with tDCS . Nonetheless, the observed changes in the white matter integrity in the left perilesional areas as well as in some right hemisphere areas present a similar pattern to changes in functional activation. Future studies with a sham-controlled arm in a larger group of patients will validate or disprove these findings (NCT01512264).In addition to functional changes in activation patterns after rTMS, white matter structural integrity may also improve. In a follow-up study,  derived from EEG can also characterize induced neuroplasticity in patients with aphasia as demonstrated by changes related to speech-language therapies  in the right hemisphere may be inefficient or deleterious to recovery. In future studies, we recommend parsing out specific functions of right hemispheric homotopic areas during spontaneous recovery, and persisting activity in some right hemispheric areas in recovery induced with NBS. Future studies should closely address the individual determinants of patterns of neuroplastic changes both to guide NBS treatment and to assess functional recovery as well as the role of neuronavigation with TMS, fMRI, PET or other techniques. In particular, future multimodal approaches pairing neuroimaging and electrophysiological measures with therapeutic NBS will more clearly define its potential in aiding rehabilitation after an aphasia-producing stroke.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Breast cancer is one of the most common cancers among the population of the Western world. Diagnostic methods include mammography, ultrasound, and magnetic resonance; meanwhile, nuclear medicine techniques have a secondary role, being useful in regional assessment and therapy followup. Optical imaging is a very promising imaging technique that uses near-infrared light to assess optical properties of tissues and is expected to play an important role in breast cancer detection. Optical breast imaging can be performed by intrinsic breast tissue contrast alone  or with the use of exogenous fluorescent probes that target specific molecules for breast cancer. Major advantages of optical imaging are that it does not use any radioactive components, very high sensitivity, relatively inexpensive, easily accessible, and the potential to be combined in a multimodal approach with other technologies such as mammography, ultrasound, MRI, and positron emission tomography. Moreover, optical imaging agents could, potentially, be used as “theranostics,” combining the process of diagnosis and therapy. Breast cancer is a major global health problem. In 2007, an estimated 1.3 million new cases of invasive breast cancer will be diagnosed and about 465,000 women are expected to die from this disease worldwide [Breast cancer is one clear example of excellent survival statistics when early-stage disease is treated using current therapies. Currently, numerous clinical methods are used in breast cancer screening and diagnosis . The mosTechniques such as magnetic resonance imaging (MRI) and ultrasound are sometimes used in addition to X-ray mammography, but have limitations such as high cost, low throughput, limited specificity (MRI), and low sensitivity (ultrasound). Thus, there is still a need to detect cancers earlier for treatment , 5, missNew methods are being investigated to bridge the current gap in clinical utility. Examples of such experimental techniques are elastography, tomosynthesis, dedicated computed tomography (CT) and positron emission tomography (PET), photoacoustic (or optoacoustic) imaging, and optical imaging.Breast imaging is largely indicated for detection, diagnosis, and clinical management of breast cancer. Commonly used imaging modalities include mammography, ultrasonography, magnetic resonance imaging (MRI), scintimammography, single photon emission computed tomography (SPECT) and positron emission tomography (PET).Mammography is, essentially, the only widely used Imaging modality for breast cancer screening. Several large randomized clinical trials have shown that mammography reduces mortality from breast cancer –12. ExteThe role of ultrasound in breast imaging has been largely limited to applications such as distinguishing between cystic versus solid masses, evaluation of palpable masses, and for needle core biopsy. In recent years, the number of indications has been greatly expanded and breast ultrasonography is now an essential modality in breast imaging. Colour Doppler and more recently sensitive power Doppler ultrasound has been used for further evaluation of the breast. In 2002, Kolb et al. published a fundamental article that showed improved sensitivity (97% versus 74%) when adjunctively used with mammography compared to physical examination with mammography . HoweverIndependent clinical trials for women at high risk of hereditary breast cancer indicate increased sensitivity with breast MRI than mammography but with variable specificity. Dynamic contrast enhanced MRI (DCE-MRI) is a very important tool for detection, diagnosis, and clinical management of breast cancer. However, it requires intravenous injection of Gadolinium-contrast agent that entails some elevated risk . Breast Radionuclide-based imaging techniques such as scintimammography, single-photon emission computed tomography (SPECT), positron emission mammography (PEM), and positron emission tomography (PET) are additional imaging techniques that provide for physiologic information. Scintimammography and SPECT typically use 99mTc-Sestamibi or 99mTc-Tetrofosmin for breast-cancer imaging. Scintimammography is used for imaging mostly palpable lesions that were occult or indeterminate from other imaging modalities. Radionuclide imaging has long been used in breast cancer management, primarily in the form of bone scintigraphy (“bone scan”) to detect bone metastases.18F-fluorodeoxyglucose (FDG) PET is increasingly used in staging advanced or recurrent breast cancer and in monitoring response to therapy; in fact, has received approval for Medicare reimbursement for these clinical indications. For detection of primary tumors, 18F-FDG-PET has been reported to have similar sensitivity as SPECT . To over There is a need to develop techniques that provide depth information in breast X-ray imaging. Currently, stereoscopic digital mammography (SDM), digital breast tomosynthesis (DBT), and dedicated breast computed tomography (BCT) are three modalities that are being actively investigated.In stereoscopic digital mammography, two projection images, spaced a few degrees apart, are acquired with a digital mammography system. A dedicated stereoscopic workstation displays orthogonally polarized images and are viewed by the observer using passive cross-polarized glasses, so that each eye visualizes one image to provide depth perception.Optical imaging techniques of the breast such as diffuse optical tomography (DOT), diffuse optical imaging, and diffuse optical spectroscopy are being investigated as an adjunct technique. Electrical impedance spectroscopy (EIS) and microwave imaging spectroscopy (MIS) are also being explored for potential use in breast cancer detection. “…a simple procedure and a valuable aid in the interpretation of pathological conditions in the mammary gland. Its use is recommended in the routine examination of the breast…” [Optical characterization of the breast has been attempted since 1929  when thebreast…” . Cutler Optical imaging as an adjunct device to mammography or ultrasound imaging is a natural choice, because the information gained with the optical signal is markedly different from that of the clinical imaging information. As a result of numerous scientific and technological advances in tissue optics since 1990, optical mammography now appears feasible with levels of specificity and resolution superior to early developments. Optical breast imaging is a novel imaging technique that uses near-infrared (NIR) light to assess optical properties of tissues, and is expected to play an important role in breast cancer detection. When fluorescent probes are excited by NIR light, they emit photons at predefined wavelength ranges, detectable by an optical imaging system.Until now, studies have focused on using the intrinsic optical properties of the breast to visualize lesions without the use of fluorescent contrast agents. These studies described higher absorption for carcinomas than for the surrounding parenchyma due to increased blood content associated with angiogenesis –25. HoweClinical optical imaging diagnosis started with the development of clinical optical breast imaging systems . CompaniMajor advantages of optical imaging are that it does not use any radioactive components (as in PET and SPECT), which can result in repeated use even in young women, and that its sensitivity is very high (nanomolar to picomolar concentration range) compared to MRI, it is relatively inexpensive, and is easily accessible. However, this technique is still in a very early phase of development. Optical imaging uses light propagation through tissue to assess its optical properties. In general, optical imaging devices transmit light through the breast, where it is both absorbed and scattered by the tissue components present. The light used in optical imaging is commonly monochromic and in the near-infrared (NIR) range permitting imaging up to several centimeters deep in soft tissue. Different tissue components have unique scattering and absorption characteristics for each wavelength. NIR in the wavelength range of 600–1000 nm is used to allow for sufficient tissue penetration. After passing through the breast, the remaining light is registered by detectors and advanced computer algorithms are used to reconstruct the images , 32. All optical imaging systems, in general, use three different illumination methods: continuous wave (CW), time-domain photon migration (TDPM), and frequency-domain photon migration (FDPM).Continuous-wave systems emit light at constant intensity or modulated at low frequencies (0.1–100 kilohertz) . The conThe time domain technique uses short (50–400 picoseconds) light pulses to assess the temporal distribution of photons . BecauseFrequency domain devices modulate the amplitude of the light that is continuously transmitted at high frequencies (10 MHz–1 GHz). Measuring photons phase-shifts and their amplitude decay , information on optical properties of tissue is acquired and scattering and absorption can be distinguished. When photon density waves encounter tissue regions of varying optical properties, they refract, scatter, and interfere. The result is a “perturbation” whose magnitude is dependent on the location, size, and optical contrast of variations within a tissue volume.During the last decade, progress in source and detector technology, light-propagation modeling, and potential fluorescent contrast agents, has resulted in a renewed interest in optical imaging .Three types of light sources are used for optical imaging applications: white light, light-emitting diodes (LED), and laser diodes. The spectrum of most white light sources extends over the visible range and into the near-infrared. In light-emitting diodes, free electrons move across a diode junction. In this process, photons are generated. The photon energy is set by the energy drop between the conduction band and the valence band. In laser diodes (or diode lasers), a population inversion is induced. Laser diodes are used widely for optical imaging applications. Photodiode is a robust and inexpensive detector for relatively high light levels. Light with energy greater than band-gap energy hits the photodiode, excites electrons into the conduction band, and creates a hole in the valence band. Avalanche photodiode (APD) is a high-speed, high-sensitive photodiode with an internal gain mechanism through a reverse-bias voltage. The electron-hole pairs are generated from exposure to light with higher photon energy than band-gap energy. Photomultiplier tube (PMT) is a sensitive detector which amplifies the input light signal 105 to 106 times with almost no additional noise. It is usually selected for high-speed or low-light level detection. It is suitable for single-photon counting applications when the rate photons striking the photocathode are below 100 MHz. Charge-coupled device (CCD) is a solid-state sensor with a wafer of silicon crystal. When the silicon is exposed to light, the photoelectric effect generates electrons from the silicon bonds. These free electrons are collected by CCD electrodes at the interface created by positive surface potential to form an extremely thin, but very dense, inversion layer. Image intensifier is a vacuum tube device consisting of a photocathode input, microchannel plate, and the phosphor screen. The electrons due to photoelectric effect are accelerated and multiplied through microchannel plate (MCP) via mechanism analogous to photomultiplier, to the phosphor screen where the light is released upon striking the coating.Optical breast imaging can be performed (1) relying on intrinsic breast tissue contrast alone ; or (2)Optical breast imaging uses NIR light to assess the optical properties of breast tissue. Light absorption at these wavelengths is minimal, allowing for sufficient tissue penetration (up to 15 cm). The main components of the breast all have specific absorption characteristics as a function of the wavelength. Imaging of scattering may be associated with structural characteristics and the concentrations of organelles. By combining images acquired at various wavelengths (spectroscopy) concentrations of oxy- and deoxy-hemoglobin, water, and lipid can be determined. In a malignant tumor, hemoglobin concentration is directly related to angiogenesis, the key factor required for tumor growth and metastases . In addiin vivo measurements show that scattering coefficients are wavelength-dependent. While ex vivo studies suggest that the scattering coefficients alone do not provide sufficient information to discriminate the small breast tumors from normal breast tissue  and the oxygen saturation (the ratio of oxygenated hemoglobin to the blood volume). Statistical data indicate that there are two-to fourfolds of contrast between normal and tumor structures for the blood volume, and the oxygen saturation in the tumor is also less than normal.A novel element that can enhance the potential applications of optical imaging is the use of contrast agents. In optical breast imaging with contrast agent, fluorescent probes are used that emit photons at predefined wavelengths after excitation by laser light. These photons are detected while the light of the excitation wavelength is filtered . FluoresLike other imaging modalities, contrast enhancement through contrast agents, either for absorption or fluorescence, have shown great promise for improving the sensitivity and specificity of breast cancer detection .The most commonly used contrast agent in the NIR spectral window is Indocyanine Green (ICG) .ICG is a nonspecific blood pool agent that is both absorbing and fluorescent in the NIR range. It is used clinically, mainly for retinal angiography and liver function tests. Both studies observed differences in ICG pharmacokinetics between malignant and benign lesions on the optical images.Important advantages of optical imaging with contrast agent are that it does not use any radioactive components (as in PET and SPECT), and that its sensitivity for probe detection is very high (possibly in the nanomolar to the 100 picomolar concentration range) as compared to MRI (micromolar to millimolar range). Moreover, optical imaging uses no ionizing radiation and can thus be used repeatedly, also in younger women. Similar to nuclear imaging, however, the optical method generally can detect very small concentrations of chromophores or fluorophores, but without using ionizing radiation and at a reduced cost.Few studies reported on optical breast imaging with the use of a fluorescent contrast agent. Two case reports described their experiences using the nonspecific agent Indocyanine Green (ICG), the only fluorescente agent approved for use in humans today –48. BothCancer cells overexpress certain receptors, and increase the uptake of the corresponding ligands. This process will result in the accumulation of those ligands in a certain type of cells, thus providing high detection specificity. Conjugation of a fluorophore to those ligands can provide high fluorescent contrast for tumor versus normal cells.Breast cancer (BC) is a heterogeneous class of disease exhibiting a variety of phenotypes and molecular profiles. For selection of the most promising therapy, with regard to the molecular profile of a cancer lesion, immunohistochemistry  is performed. In this context, identification of for example, hormone receptor positive  or human epidermal growth factor receptor 2 positive (HER-2+) cancers are important and significant points to be addressed.in vivo. A new advance that holds great promise for breast cancer research is the recent development of optical probes for molecular imaging, specifically in the NIR range. Fluorescent dyes that target specific tumor receptors , or thatin vivo and real-time imaging of the expression patterns for various enzymes, which are crucially involved in tumor formation and metastasis. Using this technology, appropriately engineered fluorescent probes can be selectively activated by endogenous or transferred gene expression. The combination of such probes with optical imaging may yield a unique, highly sensitive technology for Therefore, the impact of developing molecular-optical imaging, and in particular molecular-DOT, of the breast is potentially enormous. First, selected molecular activity can be achieved with high sensitivity. Second, cancers could be detected at their molecular onset, before anatomic changes become apparent. Therefore, therapies can be initiated at a very early stage. Third, specific cancer parameters such as growth kinetics, angiogenesis growth factors, tumor cell markers, and genetic alterations could be studied without perturbing the tumor environment. Finally, those points are important in the development of novel targeted drugs and therapies. Another exciting application of optical technology is the combination of optical imaging with other imaging modalities. Light guidance using optical fibers makes optical imaging compatible with many other radiologic methods , such asThe only two imaging techniques used, even occasionally, during oncologic surgery are X-ray fluoroscopy (for angiography) and US (for mass detection). However, the former exposes patients and professionals to ionizing radiation, and, the latter requires direct contact with tissue, and neither can be used with targeted contrast agents. Optical imaging, that exploits invisible NIR fluorescent light (700 to 900 nm), offers several advantages for image-guided surgery, including low inherent autofluorescence background, highly sensitive and specific detection of tumors up to millimetres deep in scattering tissue, and real-time imaging (reviewed in Frangioni ). IntraoOptical imaging is a very promising imaging technique. Optical imaging systems can detect imaging agents in picomolar to nanomolar concentration ranges, whereas, for instance, magnetic resonance imaging (MRI) needs larger molecular masses before probe detection is possible (at least micromolar concentrations). In contrast to X-ray, computed tomography (CT) and positron emission tomography (PET), optical imaging uses no ionizing radiation. Repeated imaging is therefore possible without radiation risks. Moreover, optical imaging is relatively cheap, and imaging agents are easy to generate and have long half-lives, particularly compared to PET tracers. Main challenges in optical imaging are depth penetration, signal quantification, and development, validation and approval of relevant imaging agents for human use. Light penetration in tissue is limited, but with the use of near-infrared light, and the development of more sensitive detection equipment, penetration in human tissue is now possible up to 15 centimeters deep. Another way to deal with the lower spatial resolution is a multimodality approach. Optical imaging could be fused with other anatomical imaging techniques, such as ultrasound (photoacoustic imaging) or MRI, similar to nuclear imaging techniques as PET/CT. The other key challenge in clinical translation of optical imaging is the development of relevant imaging agents. Crucial for this step is the full understanding of the molecular biology of breast cancer to identify potential targets. Biologic processes to study for target identification include, for example, tumor metabolism, angiogenesis, proliferation, apoptosis, and hypoxia. When clinical translation of appropriate optical imaging agents will be successful, optical breast imaging could improve early detection of breast cancer; for example, in women with dense breasts who are at increased risk for breast cancer. X-ray mammographic screening has very limited sensitivity in these women due to the tumor-hiding projection of the dense glandular tissue , 60, whi


Increasing evidence links genomic and epigenomic instability, including multiple fragile sites regions to neuropsychiatric diseases including schizophrenia and autism. Cancer is the only other disease associated with multiple fragile site regions, and genome and epigenomic instability is a characteristic of cancer. Research on cancer is far more advanced than research on neuropsychiatric disease; hence, insight into neuropsychiatric disease may be derived from cancer research results. Towards this end, this article will review the evidence linking schizophrenia and other neuropsychiatric diseases  to genomic and epigenomic instability, and fragile sites. The results of studies on genetic, epigenetic and environmental components of schizophrenia and autism point to the importance of the folate-methionine-transulfuration metabolic hub that is diseases also perturbed in cancer. The idea that the folate-methionine-transulfuration hub is important in neuropsychiatric is exciting because this hub present novel targets for drug development, suggests some drugs used in cancer may be useful in neuropsychiatric disease, and raises the possibility that nutrition interventions may influence the severity, presentation, or dynamics of disease. Genomic instability refers to an increased mutation rate that can take the form of chromosomal abnormalities, translocations, large or small insertions or deletions and base changes. Epigenomic instability refers to perturbed responses of gene regulation to environmental fluctuations. Fragile site regions of the genome have high levels of genetic and epigenetic instability.In 2003, we reported a link between somatic mutations (genomic instability) and fragile sites and schizophrenia . Later, Schizophrenia and Autism are neuropsychiatric diseases linked to multiple genetic and environmental factors. Like many common illnesses these diseases remain an enigma because there is no single factor or small number of factors that accounts for a large number of patients. 1) vary even in the same family, raising the possibility that several different diseases (i. e. the “schizophrenias”) presenting similar collections of symptoms have been grouped together )et al.  reportedo autism .The paternal age connection implicates changes to paternal germline DNA in some cases of schizophrenia because DNA is the sole paternal biological contribution to progeny. Paternal aging is linked to diminished semen quality  and fertBase substitutions account for all but progeria mutations in LMNA. The majority of mutations are transitions, (C to T) although some transversions (C to G) occur in a single dinucleotide CpG sequence. However, neither the number of replication cycles nor the observed mutation rates -113 accovia folate deficiency) in affected individuals. Generally, the specific mechanism(s) by which nutritional deficiencies produce these birth defects are unknown.Under-nutrition  and malnutrition  occur worldwide and are the most common diseases of childhood and prenatal life. Moderate to severe under-nutrition occurring prior to 2 years of age is associated with persistent behavioral and cognitive deficits that resist nutritional rehabilitation . PregnanThe importance of folic acid in preventing birth defects  is well known, although the mechanism of disease induction is not understood . Less we10). Abbreviated schemes of de novo synthetic pathways for ribo- and deoxyribo- nucleoside triphosphate synthesis are shown in Fig. (11). Folate derivatives are required by thymidine synthase that converts dUMP to dTMP, and for two steps in the purine biosynthetic pathway to make IMP; hence impacting ribo and deoxyribo purine synthesis. At the molecular level, folic acid deficiencies have the potential to disrupt nucleic acid metabolism, processes that require energy , activated nucleotide precursors , or SAM (or folate directly) for methylation . SAM is used by >100 methyl transferases that act on DNA, RNA, proteins (e. g. DNA methyl transferase DNMT (for review see [Folate participates in the methioine cycle to synthesize S-adenosyl methionine (SAM). SAM is the second most used cofactor in the cells after ATP ), histovia methyl transferases) and de-adenylated SAM.In the methionine cycle, a methyl group from folate is use by the enzyme, Methionine Synthase (MS), to convert homocysteine (HCY) to methionine. Alternatively, Betaine Homocysteine Methyl Transferase (BHMT) regenerates methionine from HCY using a methyl group from betaine (choline). Dietary and regenerated methionine reacts with ATP to generate SAM, while HCY is the product of de-methylated  HCY is up-regulated in schizophrenia patients with a 5 microM plasma HCY level associated with a ~1.7 fold increase in schizophrenia risk . 12) links DNA replication and epigenetic changes through folate and SAM metabolism, and because epigenetic marking closely follows DNA replication at the macromolecular level. HCY, a key intermediate used for SAM metabolism, is required for the synthesis of GSH; hence, dopamine metabolism, DNA replication and epigenetic marking are linked to oxidative stress.MS, the enzyme that uses folate to reform methionine from HCY, covalently adds a folate derived methyl group to the dopamine D4 receptor. The dopamine D4 receptor acts like a methyl transferase when activated by dopamine and transfers the methyl group to membrane lipid polysaccharide, changing local membrane fluidity . Dopamin9), is a common consequence of obstetric complications linked to schizophrenia [in vivo. The brain is especially sensitive to oxidative stress. Oxidative stress (hypoxia) is linked to schizophrenia directly  can impact the folate-methionine-transulfuration hub. For instance, winter births are associated with times of food scarcity [Nutrition is critical for maintaining the folate-methionine-transulfuration hub because vitamines B6, B9 (folate) and B12, and the amino acid methionine must be obtained from the diet. Other factors listed in Fig. . Reed and colleagues -146 have


The increase in the intrinsic connectivity of amygdala with the temporal poles was inversely correlated with symptom severity and anxiety scores. Although the directionality of connections between regions cannot be inferred from temporal correlations, the reduced intrinsic connectivity of the amygdala predominantly with regions involved in emotional processing may reflect impaired bottom-up signaling for top-down cortical modulation of limbic regions leading to abnormal affect regulation in MDD.Imaging studies of major depressive disorder (MDD) have demonstrated enhanced resting-state activity of the amygdala as well as exaggerated reactivity to negative emotional stimuli relative to healthy controls (HCs). However, the abnormalities in the intrinsic connectivity of the amygdala in MDD still remain unclear. As the resting-state activity and functional connectivity (RSFC) reflect fundamental brain processes, we compared the RSFC of the amygdala between unmedicated MDD patients and HCs. Seventy-four subjects, 55 adults meeting the DSM-IV criteria for MDD and 19 HCs, underwent a resting-state 3-T functional magnetic resonance imaging scan. An amygdala seed-based low frequency RSFC map for the whole brain was generated for each group. Compared with HCs, MDD patients showed a wide-spread reduction in the intrinsic connectivity of the amygdala with a variety of brain regions involved in emotional processing and regulation, including the ventrolateral prefrontal cortex, insula, caudate, middle and superior temporal regions, occipital cortex, and cerebellum, as well as increased connectivity with the bilateral temporal poles ( Major depressive disorder (MDD) is potentially a life-threatening mood disorder with substantial morbidity and mortality –3. It isA large body of evidence from neuroimaging studies suggests that depression is the result of disruptions of neural circuits encompassing large parts of prefrontal cortex (PFC), limbic, and subcortical structures , 7. The The amygdala is a critical brain region for both bottom-up and top-down processes of emotion generation and regulation . It playAs the amygdala interacts extensively within widely distributed cortical–subcortical–limbic circuits, there is increase in number of studies over the last decade examining the functional interactions between the amygdala and other brain regions to understand the disturbances in the functional connections or functional connectivity in amygdala based neural systems in depression. “Functional connectivity” refers to the temporal correlation between the neural activity patterns of anatomically separated brain regions, which is thought to reflect the level of functional communication between the regions . Most taa priori, and the resulting functional connectivity map is generated from the temporal correlations between the seed region and other brain regions  approach has been widely used in psychiatry to probe differences in neural circuits between groups without the potential confounds of task performance and practice/habituation effects. Resting-state data can be processed either by: (1) region-of-interest (ROI) or seed-based analysis, or (2) independent components analysis (ICA) . The ROI regions , 34. ICA regions .The results of resting-state fMRI studies regarding amygdala connectivity in MDD have been inconsistent. Among six resting-state fMRI studies using model-free ICA, only one study showed deceased bilateral amygdala and left insula resting-state activity and functional connectivity (RSFC) in an affective network , whereasa priori defined region such as amygdala, because of the simplicity in analysis and interpretation of the results  equipped with an eight-channel, phased-array head coil. Two resting-state fMRI scans of 230 s  in duration were acquired using a single-shot gradient-recalled echo, echo planar imaging sequence . A T1-weighted structural MRI  was also acquired for anatomical registration of the fMRI data following analysis. Images of 11 MDD patients were collected using 3 T General Electric MR scanner (Discovery MR 750) using the same parameters and protocol. Participants were required to remain still in the MRI scanner with their eyes open and fixated on a black crosshair at the center of a projection screen. The participants were instructed to relax, not think about anything in particular, and not to fall asleep.All image data were analyzed using the FMRIB Software LibrarySignal fluctuations in the white matter (WM) and cerebrospinal fluid (CSF) predominantly reflect non-neural fluctuations such as scanner instabilities, physiological artifacts, and subject motion , 56. TheZ transformation) by Student’s t-test.The left and right amygdala were first located on the standard MNI brain template, using the structures tool available in the FSLView Toolbox. Using FLIRT, these regions were then registered to each participant’s native fMRI image. Average time courses from each of the right and left ROIs were then extracted from the pre-processed resting-state fMRI. Prior to the analysis of images, the synchrony between the left and right amygdala for all participants was determined using the Pearson correlation coefficient, and was compared between MDD patients and HCs , which uses a data modeling technique based on the general linear model. The general linear model method used for first-level data  was FMRIB’s improved linear model (FILM). For the current study, a regression model was created to include the left amygdala time series as a predictor and eight nuisance covariates . The result is an estimate of how well the temporal response of the left amygdala correlates with the temporal signal of each voxel in the brain. This process was repeated again for the right amygdala.Z-statistic images were generated with a statistical threshold of Z = 2.3 (p = 0.01), and a cluster size threshold of at least 77 contiguous voxels corresponding to a corrected p-value of 0.05 as determined by Monte Carlo simulations using AlphaSimUsing FMRIB’s Local Analysis of Mixed Effects (FLAME) in FEAT, we performed higher-level analysis . Given the documented effect of age and sex on resting-state functional connectivity between regions –62, thesZ values of clusters that showed significant difference in RSFC contrast maps of MDD versus HC and each of three clinical variables . Considering the exploratory nature of the analysis, we used an uncorrected statistical threshold of p ≤ 0.05 for each test.To explore the relation between clinical variables and the abnormalities in RSFC of the amygdala in MDD, we computed the Pearson correlation coefficients between the mean Age, sex, and education were not significantly different between the MDD patient group and the comparison HC group Table . As exper = 0.87 ± 0.06; controls: r = 0.85 ± 0.11), and did not differ significantly . The RSFC analysis demonstrated significantly decreased functional connectivity of both the right and left amygdala with regions including the caudate, insula, occipital regions, and cerebellum , and HAM-A scores , in MDD patients. Furthermore, the HRSD and HAM-A scores were found to be significantly correlated  probably due to overlapping of anxiety items in both scales and psychopathology of anxiety and depressive symptoms in MDD. This might partly explain the correlation of both test scores with RSFC between left amygdala and right temporal pole. No significant correlation was found between current illness duration and left amygdala intrinsic connectivity with right temporal pole .Given that the left amygdala demonstrated reduced connectivity with more regions than the right amygdala, we performed a correlation analysis between the clinical variables and brain regions with maximum cluster size that showed reduced RSFC with left amygdala. Among the regions that showed increased amygdalar RSFC in MDD, the temporal poles were selected because of bilateral involvement. We found no correlation between clinical variables  and left amygdala connectivity with left caudate, right insula, left vlPFC, right precuneus, right cerebellum, and right occipital region. However, we found a significant inverse correlation between the left amygdala–right temporal pole RSFC and the HRSD scores  methodology to examine abnormalities in intrinsic connectivity of amygdala in patients with MDD. Confirming our hypothesis, our current data demonstrated reduced intrinsic connectivity of the amygdala with several brain regions implicated in depression including vlPFC, caudate, insula, precuneus, superior temporal gyrus (STG), occipital cortex, and cerebellum in MDD patients compared to HCs. However, we did not find abnormalities in intrinsic connectivity of the amygdala with medial prefrontal regions or ACC. Interestingly, MDD patients showed greater intrinsic connectivity between bilateral amygdala and contralateral temporal poles than healthy subjects and furthermore, intrinsic connectivity of the amygdala with right temporal pole was negatively correlated with depression severity and anxiety scores. Our results provide the first empirical evidence for wide-spread reduction in intrinsic connectivity of the amygdala with several functional networks involved in bottom-up emotional processing and regulation such as the ventral neural system , default mode network (precuneus), social cognition (STG), and visual and cerebellar regions.It is worth noting that we observed decreased intrinsic connectivity between the amygdala and the right insula using seed-based analysis, which is consistent with previous findings using an ICA approach . SimilarThe observed attenuated connectivity between the amygdala and caudate highlights the disruptions of limbic–striatal–thalamo-cortical circuits in MDD. Our results are in line with evidence from several neuroimaging studies suggesting the critical involvement of limbic–striatal–thalamo-cortical circuits in the pathophysiology of MDD , 73. BotWithin the prefrontal regions, we found decreased connectivity between amygdala and vlPFC, which has been implicated in processing emotional salience, motivation, and in the regulation of emotional and behavioral responses by modulating emotion-related brain regions such as the amygdala and ventral striatum –80. Our Contrary to previous studies, we did not find abnormalities in intrinsic connectivity between the amygdala and the medial PFC or ACC , 41. TheThe precuneus is a key part of the default mode network, a neural substrate for visual imagery and episodic memory recall, and plays a major role in self-referential processing , 83. TakThe cerebellum, although underemphasized in the neural model of depression, has been implicated in mood disorders and could play a key role in emotional perception and cognitive processing . ElectriThe amygdala has extensive reciprocal anatomical connections with the visual cortex . EmotionThe STG, temporal pole, and amygdala have been implicated in emotional processing and social cognition such as Theory of Mind , 93. TheOur results showed lateralized patterns of reduced connectivity for the left amygdala in MDD. The left amygdala demonstrated reduced connectivity with more cortical regions  than the right amygdala in MDD. These lateralized abnormalities in amygdala connectivity may reflect the structural and functional differences reported in the literature. The centromedial division of the left amygdala is 25% smaller than the right amygdala , and metThere are several limitations that should be considered when interpreting these results. First, unbalanced design may have violated the assumptions underlying group analysis especially homogeneity of variance resulting in statistical error. Hence the results need to be replicated in a balanced design. Second, interscanner and intersubject variability would have affected our results as 11 MDD subjects were scanned in a different scanner. However, we covaried for scanner differences and characteristics of MDD subjects scanned in two scanners were comparable. Third, a seed-based method limits our ability to examine functional connectivity patterns on a whole brain scale beyond the selected regions of interest. Fourth, temporal correlation analysis limits our inference on the directionality or causality of the abnormalities within the regions that are functionally connected. In order to accurately determine the directionality of altered resting-state functional connectivity of the amygdala, we must perform an effective connectivity study using methods such as Granger causality analysis –104. FurThe present study extended the findings of previous studies by investigating altered RSFC of the amygdala with the entire brain using a seed-based approach in MDD patients compared to HCs. Our study suggests abnormally decreased intrinsic connectivity between the amygdala and regions of the ventral neural system , which are known to be involved in bottom-up modulation of emotional processing and regions involved in self and social cognition (precuneus and STG) as well as posterior regions  implicated in emotional and cognitive processing. This abnormally decreased intrinsic connectivity of the amygdala could provide a neural network basis for a dysregulated, hyperactive amygdala, and for depressive symptoms including negative bias, impaired social cognition, and diminished motivational and behavioral responses in major depression.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


To date, some of the most useful and physiologically relevant neuronal cell culture systems, such as high density co-cultures of astrocytes and primary hippocampal neurons, or differentiated stem cell-derived cultures, are characterized by high cell density and partially overlapping cellular structures. Efficient analytical strategies are required to enable rapid, reliable, quantitative analysis of neuronal morphology in these valuable model systems.Here we present the development and validation of a novel bioinformatics pipeline called NeuriteQuant. This tool enables fully automated morphological analysis of large-scale image data from neuronal cultures or brain sections that display a high degree of complexity and overlap of neuronal outgrowths. It also provides an efficient web-based tool to review and evaluate the analysis process. In addition to its built-in functionality, NeuriteQuant can be readily extended based on the rich toolset offered by ImageJ and its associated community of developers. As proof of concept we performed automated screens for modulators of neuronal development in cultures of primary neurons and neuronally differentiated P19 stem cells, which demonstrated specific dose-dependent effects on neuronal morphology.NeuriteQuant is a freely available open-source tool for the automated analysis and effective review of large-scale high-content screens. It is especially well suited to quantify the effect of experimental manipulations on physiologically relevant neuronal cultures or brain sections that display a high degree of complexity and overlap among neurites or other cellular structures. High content screening (HCS) of cells based on morphological parameters is increasingly used to identify novel molecular pathways in disease or potential new therapeutic treatments. Screens targeting neuronal development or neurodegeneration in particular aim to quantify neurites (axons and dendrites). Manual analysis of neuronal morphology is time consuming and becomes impractical for large datasets. While specialized commercial software applications are available to measure neurite outgrowth, such tools are usually not openly available for user customization beyond the supplied standard interface. On the other hand, free software tools for quantitative analysis of neuronal morphology do not offer convenient automated analysis of large-scale data sets (such as those produced by genome-wide RNA interference-based screens or extensive compound library screens), and often require a significant level of user interaction ,2.Here, we describe and make freely available a bioinformatics toolkit we term "NeuriteQuant" to perform automated analysis of neurite outgrowth and branching. The toolkit is open-source and based on the free image analysis software program ImageJ. Unlike other non-commercial approaches for neuronal analysis, the NeuriteQuant pipeline provides a complete, integrated routine to facilitate genome-wide high-content analysis as well as small-scale experiments. NeuriteQuant is easily configured to process large, complex datasets produced by automated screening microscopes Figure . ResultsThe toolkit can be applied to measure neuronal differentiation, neurite outgrowth, branching and the polarization of neurites into axons and dendrites. It is optimized for images containing dozens of neurons per field of view and multiple fields per condition, so that each experimental data point represents hundreds of neurons. A key feature of NeuriteQuant is that it quantifies neurite and cell body information based on morphological criteria, not on signal intensity. The algorithm applies a series of morphological filters, culminating in quantitative measurements of neurite length, neuronal cell body area, neurite-cell body attachment points, and neurite endpoints per field. From these primary measurements, average measurements per neuron are derived for neurite length, cell body area, branch points, and neurite count.The majority of the NeuriteQuant tool is implemented as an ImageJ macro, and can be easily manipulated using a simple text editor. Reference for the ImageJ macro language is available online . AdditioA particular strength of NeuriteQuant is its flexibility due to open source implementation. This flexibility allows easy adaptation of different data sources from various imaging platforms. In addition, the toolkit is already configured with a powerful content pipeline, which facilitates streamlined management of image and annotation data Figure .In general, image data must be provided according to a naming convention that identifies individual experimental conditions . For this study we controlled a standard inverted light microscope equipped with automated filter-cube turret and automated x-y-z stage positioning using custom Metamorph journals (available upon request) to automatically generate sets of images.It is advantageous, especially for large-scale experiments, to evaluate and review automated analysis data rapidly and efficiently. NeuriteQuant fills this need by automatically creating an autonomous, web-based data browser for each analysis run , which can be used to selectively enhance either small, neurite-like structures or globular, cell body-like structures in the image Figure . First, Small structures, which usually represent debris or imaging artefacts, are excluded by rapid size filtering (step 5), which was implemented by an altered flood-fill algorithm originally included in the ImageJ package. In contrast to the standard particle analyzer built in ImageJ, this modified filter is able to filter objects enclosed by larger objects . Subsequently, a well defined, one pixel wide representation of the fibrous structure is obtained by the skeletonization function of ImageJ (step 6). Skeletonized fibrous structures that do not belong to neurites are often also found within neuronal cell bodies - therefore, the overlap between fibre structures and neuronal cell bodies is determined (step 7) and subtracted (step 8) to yield a clean, one pixel-wide representation of neurites. Due to this filter, neurites that grow on top of neuronal cell bodies are also excluded from our analysis.To identify the number of neurite endpoints, the single terminal pixel of the skeletonized fiber structures are eroded (step 9) and subtracted from the original skeleton (step 10). By subsequent subtraction (step 12) of the skeleton/cell body overlap (step 11), the neurite endpoints are derived. Neurite-cell body attachment points are identified by first applying a mask generated by dilation of the binary cell body image (step 13) to identify the proximal neurite segments (step 14). The endpoints of these proximal neurite segments are determined by erosion (step 15) and subtraction (step 16). Endpoints within the original cell body mask are removed by subtraction (step 17) to yield the majority of neurite-cell body attachment points. Since this procedure depends on the presence of endpoints in the overlap region between the dilated cell bodies and neurite skeleton, it does not identify the rare events whereby two neurites emerge very close together from a single cell body, forming V-shaped attachments. Such attachments are morphologically similar to unattached neurites that contact neuronal cell bodies tangentially, and are interpreted by our analysis as no attachment, rather than two. Our analysis also does not track neurites that grow on top of neuronal cell bodies. Thus, unattached neurites that cross neuronal cell bodies are incorrectly interpreted as two attachment points.The resulting binary images of neurites, neuronal cell bodies and neurite endpoints are quantified per field, and the following specific neuromorphological parameters are reported: total neurite length, total neuronal cell body area, average cell body cluster size, total number of cell bodies, number of neurite-cell body attachment points, and number of neurite endpoints. Quantification of the average signal intensity is also reported.If neuronal cell body overlap is negligible  the reported neuronal cell body count should provide sufficient accuracy for most applications. However, if neuronal cell bodies form higher order clusters, as is commonly observed for neurons derived from P19 cells, the average neuronal cell body number can be estimated by dividing the total neuronal cell body area by a user-defined reference size of typical neuronal cell bodies. The ratio between the average cell body structure area and the measured area of individual cell bodies can serve as an approximate measure of neuronal cell body clustering.The primary measurements generated by the NeuriteQuant tool are used to derive additional neuromorphological features, including total branch number, branch density along the neurite length, average neurite length per neuron and average length of individual neurites. For this purpose, numbers of branch points are deduced as the difference between neurite endpoints and neurite attachment points. This is valid as long as neurite and neuronal cell body detection is robust, as interrupted neurite segments or neurites that are separated from neuronal cell bodies also give rise to an increased difference between neurite endpoints and neurite attachment points. All measurements can be set up for multiple colour channels, facilitating parallel analysis of, for example, neuronal subtype morphologies or individual measurements for axons versus dendrites.post hoc data analyses.An increase in the ratio between total neurite length and neuronal cell body area can result from either increases in neurite outgrowth  or from shrinkage of neuronal cell bodies , or both in combination. In our experiments, changes in the ratio between total neurite length and neuronal cell body area usually resulted from altered neurite outgrowth. Interestingly, taxol application to primary hippocampal neurons resulted in an increase in total neurite length and an apparent decrease in neuronal cell body area (see ). The apWe first tested whether the neuronal morphology measurement algorithm in NeuriteQuant can extract neuromorphological features from differentiating mouse P19 cells, a valuable pluripotent, stem cell-like model for neuronal differentiation and neuritogenesis . These cMulticolour labelling could obviously facilitate morphological analysis of neurite length and neuronal cell body area . HoweveIn order to compare objective automatic analysis to subjective, manual tracing of neurites, we first measured neurite length both with NeuriteQuant and with the semi-automated tool NeuronJ . The NeuAs shown in Figure We next compared the performance of NeuriteQuant with other related analysis toolkits. A recently developed software tool  by Wu etPool et al  developeThe Z-factor  is a meaTaken together, NeuriteQuant trades off accuracy for speed as compared to the method of Wu et al. . In compIn comparison to freely available tools, commercial software solutions, such as HCA-Vision  can offeAnalysis speed is of particular interest, given the increasing availability of genome-wide libraries that enable functional assays amenable to high throughput automation. Therefore, as a proof of principle, we applied our automated assay in P19 cells to a focused pilot screen using RNA interference mediated gene knock-down . In thisNext we tested whether our analysis protocol is applicable to quantification of more complex morphology of primary neurons. Hippocampal neurons are a well-established model system for studying neuronal development and function . They foFigure A dose-dependent stimulatory effect of cytochalasin D on dendrite outgrowth, which is consistent with an earlier report  is clearIn agreement with previous studies ,28, we aFinally, we tested whether NeuriteQuant was compatible with analysis of neurons in brain sections. Figure In the design of NeuriteQuant, our aim was not to build a tool that corrects all potential problems that might occur during data acquisition. We rather aimed for a simple, straightforward implementation that offers robust and fast analyses. Due to this simplicity, NeuriteQuant has clearly defined limitations that need to be kept in mind concerning the interpretation of measurements:1) Image analysis is optimal at high signal-to-noise levels, which enable detection of weak neurite structures by setting low detection thresholds. Therefore, markers that are highly expressed in neurons, such as the neuronal βIII tubulin isoform (antibody TuJ1) or the dendritic marker MAP2 are preferable. Measurements via markers that stain neurites barely above noise levels are less accurate and in such suboptimal conditions, weaker neurite structures might be excluded from analysis by the thresholding procedure.2) At high cell densities, if neuronal cell bodies are not separated from each other, the neuronal cell body number can only be estimated by dividing the total neuronal cell body area by a user-defined reference size of typical neuronal cell bodies. In extreme cases  this estimation may become inaccurate. NeuriteQuant also cannot distinguish individual neurites within fasciculated bundles and reports only on the length of apparent neurite structures, whether they are made of a single or multiple, bundled neurites.3) More complex morphometric measurements, which are derived from and/or dependent on the ratios of multiple primary morphological measurements, such as the average branch density, are less accurate if only few cells or few, small neurite fragments are analyzed per field. For example, if cell viability and neurite outgrowth is drastically reduced, any falsely identified neurite segment that is disconnected from a neuronal cell body gives rise to false positive detection of neuronal branches and can therefore strongly influence the measurement of neuronal branch density, as seen in primary hippocampal neurons treated with high doses (>185 nM) of nocodazole. It is therefore imperative that such complex measurements are always interpreted in the context of successful and reliable primary measurements, and verified carefully by the user via the web-based data browser.It should be noted that many conditions that are not readily quantified by the existing NeuriteQuant software can nonetheless be identified on the basis of primary measurements. Uninterpretable images could then either be excluded, or analysis settings could be refined to extract biologically useful information. For example, large cell aggregates can be detected by measuring the average cell size. By setting an appropriate threshold, such potentially misleading images could be identified. Subsequently they could be removed if deemed an artefact, or, if they are of interest to the user, NeuriteQuant based processing could be used to capture such features for quantitative analysis.NeuriteQuant is implemented as an ImageJ macro, and therefore easily accessible for extension. Updates to the NeuriteQuant tool will be made available on the NeuriteQuant website . The curIn conclusion, we offer NeuriteQuant as a free, open-source toolkit for rapid analysis of neuronal morphology. NeuriteQuant measurements provide a meaningful characterization of neuronal morphology and they can be used to identify a wide range of morphological changes with high sensitivity. By explicitly focusing on a "per-field" analysis strategy, our method avoids ambiguities in defining single neurons and their respective neuronal arbors within a dense population of neurons, which is a typical situation for many neuronal culture systems. NeuriteQuant is especially effective in deducing neuronal parameters from relatively low-resolution images (10x) and is thus able to rapidly quantify neuronal morphology from large neuronal populations. We anticipate that NeuriteQuant will facilitate the discovery of new pathways and molecular targets in neuronal development and regeneration.NeuriteQuanthttp://www.chemie.uni-dortmund.de/groups/CB/bastiaens/dehmelt/NeuriteQuant/Windows/PC, MacOSJava/ImageJ scriptImageJ 1.38 or higher (see documentation for details)GNU GPLGNU GPL SH supervised the study. LD and SH designed and interpreted experiments and wrote the manuscript. LD conceived the analysis strategy; LD, GP, and EH performed experiments and analyzed data. All authors have read and approved the final manuscript.Text file containing a summary of the image processing algorithm and Experimental Procedures.Click here for file


Using a newly established method for the isolation of high-purity dentin matrix protein 1-positive osteocytes from bone, we have found that osteocytes express a much higher amount of RANKL and have a much greater capacity to support osteoclast formation than osteoblasts and bone marrow stromal cells. The crucial role of RANKL expressed by osteocytes was validated by the severe osteopetrotic phenotype observed in mice lacking RANKL specifically in osteocytes. Thus, we provide in vivo evidence for the key role of osteocyte-derived RANKL in bone homeostasis, establishing a molecular basis for osteocyte regulation of bone resorption.Receptor activator of NF-κB ligand , a TNF family molecule, and its receptor RANK (TNFRSF11A) are key regulators of osteoclast differentiation and function. Aberrant expression of RANKL explains why autoimmune diseases, cancers, leukemia and periodontal disease result in systemic and local bone loss. In particular, RANKL is the pathogenic factor that cause bone and cartilage destruction in arthritis. Inhibition of RANKL function by the natural decoy receptor osteoprotegerin  or anti-RANKL antibody prevents bone loss in postmenopausal osteoporosis, cancer metastases and arthritis. RANKL also regulates T cell/dendritic cell communications, dendritic cell survival and lymph node organogenesis. Intriguingly, RANKL and RANK play an essential role in the maturation of mammary glands in pregnancy and lactation. Bone homeostasis depends on the coordination of osteoclastic bone resorption and osteoblastic bone formation. We reported that RANKL induces osteoclast differentiation through activating a transcriptional programme mediated by the master transcription factor nuclear factor of activated T cells (NFAT) c1. Although it is well accepted that the RANKL-NFATc1 pathway is crucially important for osteoclast differentiation, little is known about the major cellular source of RANKL in the skeletal tissue. RANKL has been postulated to be mainly expressed by osteoblasts and bone marrow stromal cells. However, here we show that osteocytes embedded within the bone matrix are the critical source of RANKL in bone remodeling. Osteocytes, the most abundant cell type in bone, are thought to orchestrate bone homeostasis by regulating both osteoclastic bone resorption and osteoblastic bone formation, but


More than 160 Active Pharmaceutical Ingredients (APIs) have been detected in surface waters, although generally at low concentrations (ng-ug/l). Most available ecotoxicological information on these pharmaceuticals originates from acute testing using algae, Daphnia and fish; results from long-term exposures are still very scarce, particularly in vertebrates. There is, however, emerging evidence to suggest that several human and veterinary pharmaceuticals may pose a serious threat to aquatic wildlife. Still, there are only two examples where exposure to pharmaceuticals has been indisputably linked to adverse effects in wild vertebrates, i.e. the feminizing effects of the synthetic estrogen ethinylestradiol , frogs and fish (exposure via ambient water). The results show that the developing Müllerian duct (absent in fish) is a highly sensitive target for endocrine disruption and developmental toxicity in amphibians and birds.2, injected into the embryonated egg, cause a disrupted differentiation and malformations in different parts of the oviduct, including the shell gland. Egg injection of EE2 also resulted in eggshell thinning in the next generation of eggs, and a dramatically altered pattern of expression of the enzyme carbonic anhydrase in the shell gland. Eggshell thinning is probably the most serious ecotoxic effect that has afflicted avian wildlife. Based on these and other data, we propose that eggshell thinning could represent a developmental effect in the Müllerian duct rather than a direct effect of pollutants in the oviduct of the adult egg-laying bird.In birds, both diethylstilbestrol (DES) and EE2 resulted in a skewed sex ratio with almost complete feminization at an ecologically relevant concentration. A partial or complete lack of oviducts was observed in these frogs, resulting in ovulation directly into the abdominal cavity. Also the gestogenic pharmaceutical levonorgestrel (LNG), used in contraceptives and other hormone therapies, has recently been demonstrated to give rise to oviductal agenesis following developmental exposure. Unlike the impact in EE2-exposed frogs, the adult ovaries of the LNG-exposed frogs also displayed an increased fraction of immature oocytes, arrested in early meiotic prophase. We conclude that both EE2 and LNG are potent developmental toxicants at environmentally relevant exposures in frogs, giving rise to dysfunctional female sex organs with subsequent sterility.The Müllerian duct also proved to be a sensitive target of toxicity in frogs. Exposure of tadpoles to EEIn conclusion, exposure to pharmaceutical steroids during early life stages can permanently impair female reproductive organ development and fertility in lower vertebrates.


We conducted a systematic literature review with indirect comparison of studies evaluating therapeutic efficacy and toxicity associated to visceral leishmaniasis (VL) therapy among HIV infected individuals.The outcomes of interest were clinical and parasitological cure, mortality, and adverse events.PRISMA guidelines for systematic reviews and Cochrane manual were followed. Sources were MEDLINE, LILACS, EMBASE, Web of Knowledge databases and manual search of references from evaluated studies. We included all studies reporting outcomes after VL treatment, regardless of their design. Study quality was evaluated systematically by using the Newcastle-Ottawa Scale (NOS) for assessing the quality of nonrandomized studies in meta-analyses. Comprehensive Meta-Analysis software v.2.2.048 was used to perform one-group meta-analysis of study arms with the same drug to estimate global rates of success and adverse events with each drug. These estimates were used, when possible, to indirectly compare treatment options, adjusted for CD4 count. Direct comparison was pooled when available.v). The parasitological cure, an outcome that presented some degree of risk of selection and verification bias, had rates that varied widely within the same treatment arm, with high heterogeneity, hampering any formal comparison among drugs. One direct comparison of amphotericin and antimoniate was possible combining results of two studies and confirming the superiority of amphotericin.Seventeen studies reporting five treatment regimens and outcome of 920 VL episodes occurring in HIV infected individuals were included. The main outstanding difference in outcome among the treatment regimens was observed in mortality rate: it was around 3 times higher with high-dose antimony use , indirectly compared to lipid formulations of amphotericin B treatment . It was observed, also by indirect comparison, higher rates of clinical improvement in study arms using amphotericin B than in study arms using pentavalent antimonial therapy  most often results in an unfavorable response to treatment, frequent relapses, and in premature deaths. Scarce data is available regarding the treatment of leishmaniasis in HIV-infected patients (VL-HIV). Despite this, clinical decisions must be made. To aid in this task we reviewed comprehensive and systematically the available literature about efficacy and toxicity of therapeutic options for VL-HIV. PRISMA guidelines and Cochrane manual for systematic reviews were followed. Direct and indirect comparisons of nonrandomized studies were used, adjusting for CD4 count. Seventeen studies reporting five treatment regimens and outcome of 920 VL episodes occurring in HIV infected individuals were included.  Currently this option has been sidelined since more efficacious and less toxic alternatives exist In recent years, several reports have emphasized the increasing importance of visceral leishmaniasis (VL) as an opportunistic infection among HIV-positive patients in areas where both infections are endemic Leishmania/HIV co-infection, surprisingly scarce data is available regarding the treatment of leishmaniasis in HIV-infected patients. Major challenges include widespread resistance to pentavalent antimonial compounds, high treatment failure, toxicity and relapse rates Despite the prevalence, clinical implications and epidemiological impact of The review methodology followed the recommendations published by PRISMA guidelines Studies were included if reporting response to therapy to VL occurring in individuals with HIV infection. Literature search was not limited to randomized controlled trials, including all study designs. Studies involving less than ten patients; or containing a mixed population where data from HIV-infected patients or from different treatment arms could not be extracted separately were excluded. Study selection was made independently by two reviewers  and any disagreement was resolved by consensus or by discussion with a third reviewer (AR).The outcomes of interest were clinical and parasitological cure, global response , early mortality (reported death during or until 30 days after treatment), treatment interruption due to intolerance and adverse event rate and relapse rate.The selected articles were read in full to confirm eligibility and to extract data. Data extraction and quality assessment were carried out by one author and checked by a second reviewer (TOF). The following information was recorded: country; year of publication and design of the study, patient characteristics , rate of primary VL, treatment schedule, VL diagnostic criteria, VL cure criteria , loss of following, clinical and parasitological cure rate. The follow-up length and relapse rate were also recorded.The Newcastle-Ottawa Scale (NOS) 2I) statistic to evaluate heterogeneity. If significant heterogeneity was found, the results from the random effects model were emphasized and summary measures were analyzed as limited information, looking for differences in studies. Random effects model is a strategy that allows the heterogeneity inter-study would be incorporated through a broad confidence interval, generating a more conservative estimate of the measure of the effect. Clinical cure, global response and death rates analysis were performed according to the intention-to-treat analysis: the analysis was based on the total number of randomly assigned participants, irrespective of how the original study investigators analyzed the data. Publication bias was assessed by Egger's test Comprehensive Meta-Analysis software v.2.2.048 was used to perform one-group meta-analysis of study arms with the same drug to estimate pooled rates of success and adverse events with each drug. These estimates were used, when possible, to indirectly compare treatment options. These unadjusted indirect comparisons were confronted with direct comparisons, when available. Also, the indirect comparisons were adjusted for CD4 lymphocyte count by using meta-regression. Meta-regression was used to explore the relationship between event rate and CD4 count by using mixed effects regression (unrestricted maximum likelihood). We used the Inconsistency (versus low dose antimony) and thus allow direct comparison. The methodological characteristics of studies, namely inclusion and exclusion, VL disease, clinical improvement and TOC criteria are shown in Our search identified 342 articles from PubMed. EMBASE, LILACS and WEB OF KNOWLEDGE databases added 10, 2 and 3 papers respectively. After exclusions by title and abstracts, 63 potentially relevant papers were selected for full text evaluation . Two othLeishmania in samples taken in most cases from the bone marrow, spleen or liver. Few diagnoses were established by the finding of Leishmania spp. in biopsy of an unusual site such skin, tongue and gingival mucosae VL was diagnosed in 10 studies exclusively if patients had a compatible clinical illness and positive Giemsa-stained smears or culture for The efficacy of therapy was assessed by clinical and/or microbiologic criteria. The clinical response definition varied among studies. The remission of fever, improvement of hematological values and regression in the size of the spleen were the main signs observed. A complete resolution of all clinical and hematological parameters A VL episode was considered microbiologically cured when the organ used at inclusion and obtained again usually between days 25 and 45 after initiation of therapy yielded no demonstrable amastigotes by direct visualization or culture. A parasitological control study was planned for all patients in three prospective trials Five treatment regimens were reported in 17 studies included in this review. There were 13 studies with 457 VL episodes evaluating antimony compounds: five studies assessing “low antimonial dose” and eight evaluating “high antimonial dose” schedules. Therefore, we assume that a low pentavalent antimony dose was administered when the length of treatment was shorter than 28 days and/or less than 20 mg Sbv/kg/day was administered. High antimonial dose was defined as treatment with ≥20 mg Sbv/Kg/day for at least 28 days. In some studies, the antimonial treatment was combined with allopurinol, or recombinant human interferon-gamma in few patients The Newcastle-Ottawa Scale (NOS) for Assessing the Quality of Nonrandomized Studies is show in The summarized measures for initial clinical improvement, global cure and death, according to the intention-to-treat analysis, are shown in 2 0%).Clinical improvement rate using amphotericin in lipid formulation (L-Lip-AmB) was superior compared to both antimony therapy groups .The uniqv  and lipid formulation of amphotericin B  seems to be relevant, despite the overlap between the confidence intervals observed  in comparison to L-Lip-AmB  treated patients, without overlap of confidence intervals . Meta-reA variety of adverse effects were depicted, such as vomiting, diarrhea, anemia, eletrolic disturbs, pancreatic, cardiac, hepatic and renal dysfunction . The eveTen out seventeen studies reported the VL relapse rate without secondary prophylaxis and it ranged from 26 to 50%. VL relapse was diagnosed if parasites were observed in tissue samples after initial clinical cure. It was not possible to attest the presence of any difference in relapse among different treatments.v than among patients treated with amphotericin B. It could be due to the low efficacy or the toxic effects of antimony; however the risk of death seems to be related to the increase in Sbv dose, suggesting that toxicity is the most important factor. Our data confirm that antimony compounds are poorly tolerated in the presence of HIV infection, as clearly demonstrated by the study evaluating mortality among coinfected and HIV-uninfected, both treated with Sbvv than that observed with miltefosine in HIV-infected patients strongly suggests that this effect was caused by the antimonial treatment itself. Pentavalent antimonial drugs have been used for the treatment of VL since the 1940s in vivo conversion to trivalent antimony compounds may be involved in both antileishmanial activity and drug toxicity v in adults) have been used in the past v for a minimum of 28 days, which was also suggested by our data. In this review, it was also observed that, as is already accepted v, verified by an increase in the occurrence of severe events and mortality rate. Although sensitivity to specific drugs varies by region, it is unlikely that these regional differences and strains of Leishmania have contributed to the discrepancy observed in clinical response since all except one study were performed in Europe and Ethiopia, where resistance is rare. Only one study performed in India was included and, in this case, the treatment was carried out with liposomal amphotericin B and it reached a clinical response rate of 93% Available evidence suggests superiority of amphotericin B in the treatment of HIV-infected patients with visceral leishmaniasis (VL-HIV). The main conclusion of this review is the higher mortality rate among VL-HIV patients treated with SbLeishmania-HIV coinfected patients.All but one study in the L-Lip-AmB group used liposomal formulation as treatment. Liposomal amphotericin B  consists of amphotericin B packaged with cholesterol and other phospholipids within a small unilamellar liposome. The liposomal drug formulation has improved stability in blood, macrophages, and tissues, permitting more effective tissue penetration with sustained tissue drug levels, especially in the liver and spleen. This formulation has increased affinity for ergosterol and its precursors. In addition, the presence of cholesterol in the formulation minimizes interaction with mammalian cell membranes, thereby reducing toxicity The main limitation of this review is the paucity of quality evidence. On the other hand, four literature databases were searched, making it a comprehensive review. Clinical decisions must be made. To aid in this task we presented indirect comparisons, including non-randomized studies, in the same way others have done When there is no or insufficient direct evidence from randomized trials, the adjusted indirect comparison may provide useful or supplementary information on the relative efficacy of competing interventions. The validity of the adjusted indirect comparisons depends on the internal validity and similarity of the included trials Many studies used selective criteria excluding patients with more severe clinical conditions or with high risk of toxicity, such as those with renal, pancreatic or heart dysfunction. This methodological choice could have influenced toward a lower rate of adverse events and a higher percentage of therapeutic success. However, the more stringent studies were also comparative and randomized studies In all studies, the outcome ascertainment was not blinded. Indeed, either the participants or the researchers who collected disclosure information may have been aware of participants' disease status at the time of data extraction. However, most studies have clearly defined criteria for establishing cure, and in many of them parasite clearance was required. The time to cure assessment and the follow-up time was relatively uniform and adequate in all studies. So we performed the summary measures of effectiveness for each treatment regimen to perform an indirect efficacy comparison. To carry out a clinically sound analysis, we used a conservative approach and imputed outcomes for the missing and discontinued participants assuming that they did not respond to treatment. Therefore, no response includes the intrinsic lack of efficacy and toxicity limiting the completeness of the treatment. In fact, in this review there was an inverse association between adverse events rate and clinical response, as expected. Although parasitological cure rate could theoretically provide reliable information about treatment efficacy, in most studies post treatment TOC was performed only in patients with uncertain clinical response, which represents a selection bias that could underestimate response rates.In conclusion, these indirect comparisons suggest higher clinical response rate with amphotericin B than with antimony treatment, which appears to be related to less toxicity than with higher effectiveness of lipid formulations of amphotericin. Antimonial therapy carries a higher rate of drug discontinuation and a significantly higher mortality indirectly compared to treatment with amphotericin B. A relatively large body of non-comparative cohort studies supports, at this time, the use of amphotericin B as the first choice for VL treatment in HIV-infected patients. The optimal dose of amphotericin and the difference in efficacy between its various formulations remain to be established.Checklist S1PRISMA Checklist.(DOC)Click here for additional data file.


As traditional statistical methods are gradually being defeated by both the amount of data and the general absence of underlying hypotheses, data mining procedures are becoming increasingly popular and user-friendly. By combining statistical-, artificial intelligence- and database management tools, those methods are tailored for processing large quantities of information and extracting interesting patterns. Since their first application, data mining procedures have progressively been tweaked to accommodate various types of information, including social science- and biological data. However, a number of features characteristic of biological data, including high levels of measurement variability and correlation between variables, represent an additional challenge and call for specific methods. The goal of this editorial is to highlight the spatial dimension of biological data mining.The beginning of the 21Among its numerous applications, data mining plays an increasingly important role in epidemiology. In particular, it allows processing the steadily increasing volume of genomic data and helps identifying genetic risk factors. Despite ongoing progress, the mining methods currently manufactured for exploring such data still stumble over their very characteristic features and in particular their considerable complexity and diversity. Genomic data range from DNA sequences and single nucleotide polymorphisms (SNPs) to gene and protein expression levels and protein-protein interaction patterns, and further encompass structural and functional genome annotation. Accordingly, various types of data are generally treated independently and patterns emerging from any set of analyses are stitched together to form a biological answer or to generate new hypotheses.http://gis.cancer.gov/, with the Long Island Breast Cancer Study Project for instance http://li-gis.cancer.gov/, has a long-lasting research program investigating geographic and environmental patterns of cancer using GIS technologies. Other initiatives such as cancer atlases (e.g. http://www.cdc.gov/Features/CancerAtlas/) testify to the utility of spatial analyses in epidemiology.Occasionally, such patterns are projected onto a geographical map, superimposed to migration patterns or correlated to environmental factors, placing crude numeric information into a spatio-temporal perspective [reviewed in ]. IntegrGIS-based approaches have so far proven useful in retrospectively identifying spatial diseases patterns or environmental factors likely to contribute to the expression of genetically determined diseases. However, GIS technologies are still underexploited and underrated in biomedical sciences. Here we suggest that there is room for creative minds to imagine innovative ways to apply these techniques and to merge them with data mining procedures. For example, prior to engaging into a genome-wide association study and testing vast numbers of SNPs independently for a correlation with disease susceptibility, geo-referenced population-wide SNP data could be superimposed to maps of disease rates to identify subsets of candidate disease-associated SNPs. Subsequent case-control studies could then be performed on those subsets to corroborate or refute the hypothesis of a true association. The spatial dimension of biological data in epidemiology and other areas such as image analysis provides unique challenges and opportunities for biological data mining.


The most potent malaria vectors rely heavily upon human blood so they are vulnerable to attack with insecticide-treated nets (ITNs) and indoor residual spraying (IRS) within houses. Mosquito taxa that can avoid feeding or resting indoors, or by obtaining blood from animals, mediate a growing proportion of the dwindling transmission that persists as ITNs and IRS are scaled up.Increasing frequency of behavioural evasion traits within persisting residual vector systems usually reflect the successful suppression of the most potent and vulnerable vector taxa by IRS or ITNs, rather than their failure. Many of the commonly observed changes in mosquito behavioural patterns following intervention scale-up may well be explained by modified taxonomic composition and expression of phenotypically plastic behavioural preferences, rather than altered innate preferences of individuals or populations.Detailed review of the contemporary evidence base does not yet provide any clear-cut example of true behavioural resistance and is, therefore, consistent with the hypothesis presented.resilient, rather than resistant. Ongoing national or regional entomological monitoring surveys of physiological susceptibility to insecticides should be supplemented with biologically and epidemiologically meaningfully estimates of malaria vector population dynamics and the behavioural phenotypes that determine intervention impact, in order to design, select, evaluate and optimize the implementation of vector control measures.Caution should be exercised before over-interpreting most existing reports of increased frequency of behavioural traits which enable mosquitoes to evade fatal contact with insecticides: this may simply be the result of suppressing the most behaviourally vulnerable of the vector taxa that constituted the original transmission system. Mosquito taxa which have always exhibited such evasive traits may be more accurately described as behaviourally  Anopheles funestus, which feeds almost exclusively upon human blood and predominantly feeds and rests indoors . T. TAn. arThese simulations illustrate how reduced frequencies of vulnerable traits among vector mosquitoes in residual transmission systems may, counter-intuitively, reflect intervention success rather than failure Figures  and 2. BAn. arabiensis in particular, and none of these models assume any genetic adaptation of the vector population through heritable alterations of host preference.Consistent with several other contemporary theoretical studies ,53, all In order to test this hypothesis, the existing evidence base was reviewed to identify any unambiguous examples of altered frequency of innate behavioural preferences of taxonomically homogenous wild malaria vector populations following IRS or ITN scale-up.An. gambiae complex reported from Tanzania can probably be attributed to the apparent selective suppression of An. gambiae by ITNs, leaving a transmission system dominated by An. arabiensis. Similarly, the An. gambiae population in Bioko Island [An. gambiae complex which were caught resting or feeding indoors, as well as their mean biting time [An. gambiae and An. arabiensis, making the latter less vulnerable to control with IRS [An. arabiensis can also adeptly enter and leave houses without exposing themselves to IRS or ITN formulations of pesticides to which they are fully physiologically susceptible [An. nuneztovari, An. darlingi and Anopheles punctimacula in the Americas during the GMEP [An. farauti in the Solomon Islands [Several reports of apparent change in mosquito behaviour can readily be explained by changes in species composition of the vector population, rather than any heritable modifications of the handful the taxa that contribute to persisting transmission. For example , the chao Island , was orio Island  and theno Island . Lookinging time . These bwith IRS . Recent ceptible ,48. Thisceptible  and so cceptible ,56. It ithe GMEP . The onlthe GMEP  relates  Islands . HoweverA worrying recent study in Benin reported apparently negligible impact upon malaria transmission by universal coverage schemes for pyrethroid-based ITNs, as well as their supplementation with carbamate-based IRS and insecticide-treated wall linings (ITWL) relative to a reference group of villages receiving only targeted coverage with ITNs . AlthougA more worrying recent report from Senegal does raise strong, substantive concerns about the weak impact of vector control, and even rebounding mosquito populations, associated with behavioural and physiological resistance . While iTo conclude, it is clear that intervention-mediated selection for behavioural resistance in the strict sense, meaning an increase in the frequency of heritable behaviour traits in taxonomically homogenous populations which enable them to evade fatal contact with insecticides ,56, is oNo entomological survey can measure the physiological or behavioural characteristics of dead mosquitoes that have been removed by successful intervention programmes Caution should therefore be exercised before over-interpreting most existing reports of increased frequency of behavioural and physiological resistance traits: this may simply be the result of suppressing the most physiologically susceptible and behaviourally vulnerable of the vector taxa that constituted the original transmission system. Furthermore, none of these field studies can unambiguously attribute these observations to altered frequencies of heritable behavioural preference traits, rather than altered expression of phenotypically plastic behavioural traits in an environment that has been changed by intervention coverage. The importance of plasticity in anthropophagic, endophagic and endophilic behavioural preferences in stabilizing malaria transmission against intervention efforts has long been appreciated ,17,33,60Delays in malaria eradication programmes are caused more by non-response of fully susceptible vectors to attack measures than by physiological resistance, though the latter receives more attention[.attention.modification, adapt, shift and resistance in relation to reports of apparent changes in mosquito behaviours. The term resilience, as applied to humans [resistance which infers increasingly ability to do so [Greater terminological caution is therefore warranted in relation to use of the terms o humans -63 and eo humans  may, theto do so ,56.An. funestus and malaria transmission in South Africa was clearly associated with emergence of physiological resistance to pyrethroids [Although the contributions of behavioural and physiological resistance to apparent vector population rebound in Senegal remain unclear, there is no reason to doubt the evidence  that thiethroids , but wasethroids . Both exethroids -69. In tethroids  have illethroids , can accethroids ,32,65.Ongoing national or regional entomological monitoring surveys of physiological susceptibility ,71 shoulWhile such entomological parameters can be monitored prospectively, they can only be used to infer the suppression or rebound of malaria vectors and transmission where appropriate retrospective baseline data are also available ,32. SuchA suggested generic plan for strengthening national or regional malaria vector monitoring platforms to incorporate assessment of essential behavioural phenotypes and their influence upon vector control impact, mosquito population dynamics and epidemiological outcomes.1. Expand and consolidate any existing national network of sentinel surveillance sites for physiological resistance of malaria vector mosquitoes to insecticides, ideally integrating with similar platforms for other common mosquito-borne diseases, such as lymphatic filariasis. Such sites should also overlap both with existing historical entomological study sites for which baseline legacy data is available, and with national platforms for assessing malaria burden through cross-sectional malaria indicator surveys or quality-assured facility-based surveillance.2. Establish an affordable, practical longitudinal community-based (CB) mosquito trapping scheme  with a s3. Given the reliance of scalable CB trapping schemes upon essentially unsupervised field-4. based personnel, it is also essential establish a quality assurance system in which each of these sites is regularly and randomly re-surveyed by a centrally coordinated, specialist entomological team using the same trapping methods. It is essential that the CB personnel are unaware of the re-survey schedule so that the quality of CB sampling assessed is representative of that implemented all year round.5. Establish experimental hut capacity at one or two of these sentinel sites, chosen so that most nationally-relevant or regionally-relevant vector species are available at useful densities for as much of the year as possible, enabling the efficacy of vector control interventions to be assessed before and after their introduction .6. Incorporate surveys of vector feeding and resting behaviours, using human landing catch by participants protected with drug chemoprophylaxis  and back7. Integrate questions relating to relevant human behaviours , vector CB: Community-based; GMEP: Global malaria eradication programme; IRS: Indoor residual spraying; ITN: Insecticide-treated nets; ITWL: Insecticide-treated wall lining.The authors declare that they have no competing interests.NJG and GFK conceived this analysis and review. NJG conducted literature search, drafted the manuscript and edited it based on comments from GFK. PPC and GFK contributed strategies for practical monitoring of mosquito population dynamics on national scales. GFK formulated and implemented the model simulations. All authors agreed to the final version.


Chronic disorders, such as rheumatic disease, impact children emotionally, socially and physically. Children who perceive their medical condition interferes with the ability to engage in social, educational and family activities report more depressive symptoms. The purpose of this study is to examine the prevalence of depression in children with rheumatic disease and to identify concomitant risk factors for depression in this population. It was hypothesized that, compared to healthy peers, children with rheumatic diseases may have a higher risk of developing depressive symptoms.Participants were children, 8-17 years old, followed by a pediatric rheumatology practice and identified at a routine follow-up appointment. Children completed the Childhood Depression Inventory (CDI) and parent-child pairs completed a series of standardized measures documenting demographics, symptoms, health-related quality of life, coping mechanisms and psychosocial variables. The primary rheumatologist documented a global assessment of disease severity as well as disease activity scores for children with the diagnosis of juvenile idiopathic arthritis (JIA), juvenile dermatomyositis (JDMS) or systemic lupus erythematosus (SLE). One-way ANOVAs were completed to determine if demographic variables were significantly associated with depression. Pearson product moment correlations were run to determine relationships between the depression, pain, coping, quality of life, family conflict, physical functioning and disease severity. Hierarchal multiple regression analyses were run to determine which medical and psychosocial variables were significant predictors of depression in children with rheumatic disease, while controlling for demographic variables.At this time, 96 patients have been screened  and only data regarding demographics, length of diagnosis and physician documented disease activity has been examined. Five percent of the population was found to have clinically significant symptoms of depression while 15% of the population was found to be “at risk” for symptoms of depression. Seven percent of participants were receiving medication for depression and an additional 7% were involved in therapy at screening. Medication usage significantly correlated with increased CDI scores (increased depressive symptoms) whereas those in therapy had CDI scores comparable to their non-depressed peers. Total CDI score also significantly correlated with female gender and an increased number of children in the home. Neither years since diagnosis, active joint count, total SLEDAI score, JDMS disease activity score nor physician global assessment correlated with increased CDI scores. Additional analyses of psychosocial factors are in progress.The prevalence and risk of depression among children with rheumatic diseases is comparable to previously reported data for a healthy population of children. Female gender and number of siblings are associated with increased risk of depressive symptoms. Early data suggests that medications appear to be less efficacious at controlling depressive symptoms than intervention with therapy.Stacey E. Tarvin: None; Nicole M. Taylor: None; Christine Raches: None; Lisa Macharoni: None.