116 There are several strategies to use exosomes as a (therapeutic) vaccine. Tumor-derived exosomes carrying tumor antigens and plasmacytoma cell-derived exosomes may be used to induce tumor-specific immunity and thus to prevent tumor development.117 Despite the extensive studies on EVs, until now there are no protocols available for standardized collection, isolation and storage of EVs. Such standardized protocols are important to be able to compare results between laboratories. Despite the fact that blood is probably our most complex body fluid, EVs present in

or isolated from blood or fractions thereof have been most extensively studied so far. Although there are several recommendations BAY 80-6946 cost regarding the collection of blood with regard to EVs,118 for other body fluids no protocols are available. In most studies EVs have been isolated from body fluids by differential centrifugation.[3] and [47] Differential centrifugation involves multiple sequential centrifugation steps where in each step the centrifugal force is increased to separate smaller and less dense components from the previous step. Another type of separation by means of centrifugation

Idelalisib is density-gradient ultracentrifugation, which separates vesicles based on density.[20] and [119] Although different types of vesicles have been distinguished based on density,[3], [20] and [41] differences in density are likely too small to allow full separation of EV species. Differential centrifugation and density-gradient centrifugation protocols

are unlikely to Montelukast Sodium isolate only a single type of vesicle. Immunoaffinity-based assays, usually coated with a specific CD-antibody, are also used.[84] and [120] Theoretically, this method isolates only one subpopulation of vesicles. Unfortunately, in daily practice successful isolation and purification of a single population with an acceptable recovery by this technique are usually very difficult. Ideally, EVs are measured directly in freshly collected samples, but in a clinical setting this is hardly feasible at present. When samples are frozen and thawed before analysis, concentrations and exposure of PS can markedly increase in samples containing PMVs.[35] and [118] As EVs may expose one or more surface antigens of their parent cell, the cellular origin of EVs can be assessed by using antibodies directed against such cell-type specific surface antigens. Flow cytometry (FCM) is still commonly used to estimate the number of EVs. Due to the fact that the refractive index of vesicles is low, only the larger vesicles will be detected as single vesicles and the smaller vesicles will be detected only as a swarm.121 Thus, FCM will underestimate the number and concentration of vesicles. Although many researchers use annexin V to identify or isolate MVs, PS exposure by MVs is still ambiguous because exposure of PS can be due to isolation and handling procedures such as centrifugation and storage.

Orlandini for animal care, Henrique B. Biehl and Carlos E. L. Santos for their assistance with confocal microscopy (Centro de Microscopia Eletrônica da UFRGS). Technical assistance

was provided by Silvia Barbosa and Antônio Severino. We also thank Ana Paula Horn and Lauren Valentim for their helpful information in immunofluorescence. This study was supported by grants from CNPq and CAPES. There was no conflict of interests. “
“For the growing number of people who have the risk of, or experienced cerebral infarction or TIA (Weimar et al., 2010 and Hata et al., 2005), development of a novel compound to protect neurons from LY2109761 in vivo focal ischemia, or even to promote cerebral repair, is urgently required. In the incretin family, glucagon-like peptide-1 (GLP-1), or insulinotropic Navitoclax chemical structure secreted from L cells in the gastrointestinal tract as a response to food ingestion (Cefalu, 2010 and Rizzo et al., 2009), acts as a trophic factor for β cells in the islets by enhancing insulin biosynthesis/release and their proliferation ( Turton et al., 1996). In addition to the β cell-trophic/insulinotropic

effect, GLP-1 exerts a neurotrophic effect in the brain ( McClean et al., 2010 and Perry et al., 2002). Indeed, GLP-1 can enter the brain; the GLP-1 receptors (GLP-1R) is expressed widely in the central nervous system ( During et al., 2003 and Turton et al., 1996); and the activation of GLP-1R was found to improve cognitive performance ( Li et al., 2010a and During et al., 2003). However, once secreted Forskolin mw into the blood, GLP-1 is rapidly degraded and inactivated following release

of the intrinsic antagonizing enzyme, dipeptidylpeptidase-4 (DPP-4). Exendin-4 (Ex-4), a long-acting analog of GLP-1 (a GLP-R agonist), developed for intravenous treatment of type II diabetes mellitus (DM-2), demonstrated a neuroprotective property in vivo after cerebro-ventricular administration (Li et al., 2009). Ex-4 also exerted a neurotrophic property in vitro (Li et al., 2010c). Moreover, in a transgenic mouse model of Alzheimer’s disease (AD) combined with streptozocin-induced DM-2, a continuous subcutaneous injection of Ex-4 reduced the levels of amyloid-β (Aβ) protein in the brain ( Li et al., 2010b). Alogliptin benzoate (AGL), a potent and highly selective inhibitor of DPP-4, developed for once-daily oral treatment of DM-2, demonstrated a lower incidence of unfavorable side effects such as hypoglycemia and hyperphagia, compared to previous drugs (Moritoh et al., 2008 and Feng et al., 2007). Although treatment with AGL for a prolonged period in DM-2 patients is expected to protect β cells and prevent atherosclerotic vascular damage, it is unknown whether AGL, independent of its insulinotropic properties, protects neurons against lethal ischemia.

2B). In response to M. tb antigen stimulation, QFT-IT plasma IFN-γ, IL-2, and CXCL10 responses were significantly higher in active TB and LTBI groups than in the control group (P < 0.01, Fig. 3A). TB patients also presented higher levels of IL-13 than did the control group although the differences were not significant (P > 0.05). QFT-IT plasma VEGF-A did not differentiate between active TB and LTBI groups unlike serum VEGF-A, and none of the 17 analytes differed between the two groups in

response to M. tb antigens ( Fig. 3A). All cytokines were highly produced in response to mitogen (PHA) without any significant difference between the groups (P > 0.05), suggesting that there were no non-specific immunosuppression effects on the cytokine responses to M. tb antigens BIBF-1120 in the QFT-IT plasma samples ( Fig. 3B). The effect of anti-TB treatment on immune responses was monitored 2 and 6 months after the initiation of anti-TB treatment. In the sera from TB patients, the sCD40L concentration significantly increased along with M. tb clearance in culture at the 2-month

evaluation (P < 0.001, Fig. 4). Increased serum sCD40L concentrations were present in 79% (30 out of 38) of TB patients after 2 and 6 months of treatment. Ribociclib One out of 38 patients at pre-treatment and 6 months post treatment did not have positive sCD40L concentration while all of the 38 patients showed positive sCD40L concentrations (>110 pg/mL) after 2 months of anti-TB treatment ( Supplementary Fig. 2). The proportion of the responders who showed <7000 pg/mL of serum sCD40L at baseline (59.5%; 22 out of 37) was reduced to 18.4% (7 out of 38) and 18.9% (7 out of 37) after 2 and 6 months of treatment, respectively ( Supplementary Fig. 2). Meanwhile, the number of TB patients showing >7000 pg/mL of sCD40L increased from 16 (43.2%) to 32 (86.5%) following anti-TB treatment ( Supplementary Fig. 2).

Serum VEGF-A concentrations were reduced in Arachidonate 15-lipoxygenase more than half of TB patients (55.3%; 21 out of 38) after 6 months of treatment, whereas the change in median concentrations between pre- and post-treatment was not statistically significant (P > 0.05). Sera concentrations of the other analytes, including IFN-γ, did not change during anti-TB treatment in 38 TB patients ( Fig. 4). In the QFT-IT plasma obtained from active TB patients, the IFN-γ responses were dramatically decreased in 85.7% (12 out of 14) of the TB patients after 2 months of treatment. Eight out of the 12 patients showed confirmed M. tb in culture at diagnosis while M. tb clearance was observed along with the reduced IFN-γ responses at 2 months post treatment. Additionally, all patients showed reduced IFN-γ responses post-treatment (P < 0.001, Fig. 5). Eight out of 14 TB patients showed positive TNF-α responses at baseline and the TNF-α responses decreased in all of the responders after 2 months of treatment (P < 0.05, Fig. 5). Furthermore, 69.2% (9 out of 13) and 58.

Especial agradecimento ao Dr. Mário Silva (do serviço de Anatomia Patológica dos HUC) pela sua disponibilidade na cedência das fotografias do exame histológico e na interpretação das mesmas, e

a Dra. Catarina Fontes CAL-101 concentration (do serviço de Radiologia dos HUC) pelo interesse no caso clínico. “
“O vírus da hepatite D (VHD) pertence à família Deltaviridae e é o único vírus animal satélite conhecido 1. Foi descoberto em 1977 por Mario Rizzetto et al., em Itália 2. É um vírus de ARN que necessita da presença do vírus da hepatite B (VHB) para completar o seu ciclo biológico, pelo que a infecção pelo VHD ocorre apenas em doentes infectados pelo VHB1 and 3. O seu genoma, o mais pequeno do reino animal, contém apenas 1700 nucleótidos, sendo constituído por um ARN circular, que codifica uma proteína estrutural que é o antigénio find more (Ag) delta2 and 3. O virião do VHD consiste num complexo formado pelo ARN-VHD e o Ag delta, protegidos por um envelope proteico constituído pelo AgHbs. O AgHBs é necessário para a transmissão e replicação do VHD que ocorre exclusivamente no

núcleo dos hepatócitos. Estão identificados 8 genótipos do VHD, cada um com curso clínico e localizações geográficas características4 and 5. Estima-se que mundialmente 15 a 20 milhões de doentes estejam infectados pelo VHD, correspondendo a 5% dos doentes com infecção crónica pelo VHB3. O vírus partilha as vias de transmissão associadas ao VHB, nomeadamente parentérica, sexual e intrafamiliar2. O VHD é transmitido apenas a indivíduos com infecção pelo VHB, podendo ocorrer em doentes com infecção crónica prévia pelo VHB (superinfecção) ou ser adquirido concomitantemente aquando da infecção aguda pelo VHB (coinfecção). No primeiro caso, pode manifestar-se com quadros de exacerbação de doença estável e possui habitualmente caráter dominante e de repressão sobre o VHB. O diagnóstico baseia-se no doseamento dos marcadores serológicos e da carga viral de ambos os vírus 1 and 2. Doentes com doença hepática crónica VHD-VHB têm indicação para tratamento, devendo este ser dirigido ao vírus dominante3 and 4. Apresentamos um doente do sexo masculino, 42 anos de idade, natural

da Moldávia, residente em Portugal, desde 2001. Assintomático, referenciado à consulta de Hepatologia em fevereiro 2005 por infecção crónica VHB, conhecida desde os 28 anos de Phosphoglycerate kinase idade, sem sintomas na altura do diagnóstico. Referia relações sexuais não protegidas, mas negava o consumo de drogas endovenosas, transfusões sangue, tatuagens ou piercings. Referia abstinência alcoólica, no último ano, e consumo inferior a 30 g/dia, nos 15 anos anteriores. Desconhecia a existência de doença hepática em qualquer familiar. O exame objetivo não mostrava alterações. Analiticamente, verificou-se elevação das aminotransferases (ALT 107 UI/L; valor de referência (VR) < 41 UI/L) e confirmou-se a presença de infecção pelo VHB: AgHBs, AcHBc total e AcHBe positivos e AcHBc IgM e AgHBe negativos, apresentando ADN-VHB igual a 1,8 × 103 UI/mL.

Currently, it is not clear how the sensory loss from the anterior two third of tongue alters serotonergic neurotransmission in the hippocampus. The peripheral gustatory system consists of the neural–epithelial Seliciclib clinical trial machinery linking the sensory

epithelial cells in the oral cavity to the first gustatory relay centre in the brain. Branches of the facial and glossopharyngeal nerves, which synapse with receptor cells in the taste buds, convey taste messages to the first relay nucleus, the rostral part of the nucleus tractus solitarius in the medulla.30 Taste information reached taste neurons in the nucleus tractus solitarius is relayed to other brain regions such as the hypothalamus, the ventral tegmental area and the nucleus accumbens via the parabrachial nucleus.2 and 17 In humans, striatal dopamine release reflects the perceived pleasantness of a meal.3 Intra-oral infusions of sweet and bitter stimuli learn more differentially modulate dopaminergic activity in the nucleus accumbens.31 Taste of aversive flavour increased serotonin release in the hypothalamus.32 These reports together suggest that long-term disruptions in taste sensation may reduce dopaminergic and/or serotonergic activities

in the brain regions. Sensory deprivation with bilateral olfactory bulbectomy, a well-known animal model of depression, results in a complex constellation of behavioural, neurochemical, neuroendocrine, and neuroimmune alterations.33 Especially, serotonin neurotransmission was decreased

in the hippocampus of olfactory bulbectomized rats.34 Morales-Medina et al.,35 have suggested that the lack of input from the olfactory bulbs may result in serial neuronal rearrangements in the piriform cortex, entorhinal cortex and hippocampus leading, at least partially, to behavioural deficits in emotion process. In the same study, dendritic structures in the nucleus accumbens were not affected by olfactory bulbectomy.35 Autophagy activator Together with the present study, we propose that the hippocampal dysfunction such as decreased serotonin neurotransmission is a common mechanism involved in the pathophysiology of depression by sensory deprivation in taste or olfaction, and serotonergic activity or dendritic structures in the nucleus accumbens may not play a key role in it. All listed authors have no conflict of interest to disclose. Funding was provided by Seoul National University Dental Hospital (SNUDH) Research Fund (grant #02-2012-0001). Approved by Seoul National University Institutional Animal Care and Use Committee SNUIACUC Approval No.: SNU-120725-3-2. Authors JWJ and JHL designed the study and wrote the protocol. Authors YJC, JYK, WPJ and YTK performed the experiments. Authors JWJ, YJC, JYK and YTK managed the literature searches and analyses, undertook the statistical analysis. Author JWJ and YJC wrote the first draft of the manuscript. All authors contributed to and have approved the final manuscript.

Patients were recruited according to the updated diagnostic criteria of IIH and papilledema was documented in all subjects by an ophthalmological examination including funduscopy. Twenty-five individuals with other neurological disorders served as controls. Sonographic evaluation of the optic nerve was possible in all participants. Compared to controls the ONSD was significantly enlarged among patients with IIH bilaterally [6.4 ± 0.6 mm vs. 5.4 ± 0.5 mm].

After lumbar puncture with a therapeutic removal of 30–50 ml of CSF we observed a significant decrease of the ONSD on both sides (right ONSD 5.8 ± 0.7 mm, left ONSD 5.9 ± 0.7 mm) (Fig. 1). However, in some patients with IIH, the ONSD was not altered or only slightly altered, e.g. a decline of 0.4 mm or more was only documented in five buy Enzalutamide individuals. This may possibly be related to findings of a defective CSF circulation in the optic nerve sheath in this disorder, a state that is referred to as optic nerve compartment syndrome [23]. The ROC curve analysis revealed an optimal

cut-off value for predicting raised ICP of 5.8 mm with a sensitivity of 90% and a specificity of 84%. The mean optic disk elevation in subjects with IIH was 1.2 ± 0.3 mm. Nevertheless, one patient showed no evidence of optic disk elevation in transbulbar sonography but had signs of papilledema in funduscopy. Corresponding to previous studies, we found no decrease of the optic disk elevation after lumbar puncture in the observation period

of 24 h. As a result sonographic ONSD evaluation may be useful in detecting raised ICP in patients Smad inhibitor with presumed IIH. Furthermore, our data suggest a potential usefulness of this technique for monitoring of treatment effects. In addition, ONSD values and optic disk levels were slightly asymmetric, reflecting the complex anatomy of the subarachnoidal space of the optic nerve and its possible influence on the cerebrospinal fluid dynamics. For this reason we recommend that each eye should be evaluated separately and mean ONSD values should be designated for both eyes. Predominantly, the relationship of ONSD alterations and ICP changes was verified in clinical situations with raised ICP. One case series and one prospective study investigated the ONSD in spontaneous intracranial hypotension [24] and [25]. C59 cell line Examining the orbit with T2-weighted MRI techniques, they observed a collapsed optic nerve sheath. Dubost et al. published an ultrasound study on ten patients with postdural puncture headache after lumbar puncture or epidural anesthesia [26]. Consistent with the mentioned MRI-results a small ONSD of 4.8 mm was detected before treatment. After successful therapy with a lumbar epidural blood patch a marked enlargement of the ONSD was found. Accordingly, in one patient in whom the intervention failed to resolve the headache they recorded no ONSD distension.

The Ishikawa cells were purchased from the American Type Culture Collection (Manassas, VA) and were passaged in our laboratory for less than 6 months. Cells were grown in Dulbecco’s Modified Eagle’s Medium supplemented with glutamine, pyruvate, antibiotics, and 10% fetal calf serum in a humidified atmosphere containing 5% CO2 at 37°C. Cell lysate proteins Selleck GDC-0980 were separated

by sodium dodecyl sulfate–polyacrylamide gel electrophoresis (10% gels) and transferred to nitro- cellulose membranes. Protein amounts were quantified using the Bradford method, and equal protein amounts were loaded to the gel. Membranes were blocked in TBS with 0.05% Tween 20 (TBST) containing 5% nonfat dry milk powder for 1 hour. Western blots were probed with primary antibodies for 1 hour, washed three times with TBST, and then incubated with the appropriate secondary anti- bodies for 30 minutes. Membranes were then washed with TBST three times before

developing with SuperSignal West Dura chemi-luminescent substrate (Pierce, Rockford, IL). The comet assay used to measure DNA damage has been described previously [15]. Briefly, cells were treated with 20 μM etoposide (Sigma, St Louis, MO) for 4 hours, and the damage distribution was measured as tail moment (product of tail length and fraction of DNA). Cells were harvested and resuspended in Hank’s Balanced Salt Solution (Sigma) with 10% DMSO and 0.5 M EDTA. The cell suspension was then suspended in 0.7% low-melting agarose at 37°C (Sigma) and layered on to comet slides (Trevigen, Gaithersburg, selleck chemicals llc MD). The cells were then lysed in lysis solution containing 2.5 M NaCl, 100 mM pH 8.0 EDTA, 10 mM Tris-HCl, 1% Triton X (Sigma) at 4°C for 1 hour. Denaturation was carried

out for 40 minutes, in chilled alkaline elec- trophoresis buffer (pH 13.0-13.7). Electrophoresis was subsequently carried out for 20 minutes. Slides were immersed in neutralization buffer (500 mM Tris-HCl, AMP deaminase pH 7.4), dehydrated, dried and stained with SYBR Green dye (Invitrogen, Carlsbad, CA), and scored with OpenComet plugin of ImageJ software. The images were captured using fluorescence microscope (Carl Zeiss, Oberkochen, Germany) equipped with triple-band filter. Fifty comets per sample were randomly selected and analyzed. The extent of DNA damage was expressed as tail moment, which corresponded to the fraction of the DNA in the tail of the comet. Briefly, male BALB/c athymic nude mice (4-5 weeks old) were obtained from the Experimental Animal Center of Shanghai Insti- tutes for Biological Sciences (Shanghai, China). Mice were randomly divided into the following two groups: nonsense group and missense group (15 mice per group). Nonsense-group mice were injected sub- cutaneously into the right flank with 1.0 × 107 Ishikawa cells stably transfected with PTEN nonsense mutant (R130*), whereas the missense-group mice were injected with 1.

This finding was confirmed a year later on larger number of patients in the study which compared echogenicity of the BR between 40 patients with unipolar depression, 40

patients with bipolar disorder and 40 healthy controls. PI3K assay Raphe echogenicity in patients with unipolar depression was found to be distinctly reduced as compared with healthy adults and patients with bipolar affective disorder. BR echogenicity, on average, was halved in the unipolar depressed group. No correlation was found between BR echogenicity and age, sex or disease severity [3]. Reduced brainstem midline echogenicity of depressed patients was interpreted as a structural alteration of the dorsal raphe nucleus or fiber tracts in this region [14]. Increased T2-relaxation time in a pontine brainstem in patients with major depression could be in line with previous

reports of brainstem pathology in these patients [14]. The observation might indicate a subtle tissue alteration, which cannot be identified by visual inspection of the images. T2-relaxation time depends on physical tissue characteristics and is influenced by hydration status or iron content. Differences in T2-relaxation time of specific brain areas between patients with major depression and healthy controls may indicate different tissue composition caused by histological check details changes. Several further studies confirmed the finding of reduced echogenicity of the BR in unipolar depression. In the study Glutathione peroxidase of Walter [17] the frequency of patients with reduced echogenicity of BR was higher in unipolar depression compared with healthy individuals and in depressed PD patients compared with non-depressed. The

frequency of reduced echogenicity of BR was the highest in patients with unipolar depression. In this study, reduced echogenicity of the BR was more frequent in depressed than in non-depressed patients, irrespective of presence of PD. TCS findings of another study [19], showed that reduced echogenicity of pontomesencephalic BR is frequent in depressive states, irrespective of diagnostic category of depression, but only rare in healthy subjects without any history of psychiatric disorder. BR echogenicity could not discriminate between major depressive disorder and adjustment disorder with depressed mood. BR echogenicity scores showed in this study were significantly lower in SSRI responders compared with SSRI non-responders. Reduced BR echogenicity indicated SSRI responsivity with a positive predictive value of 88%. Recently, reduced raphe echogenicity was found in 47% of the patients with major depressive disorder but only in 15% of healthy controls. In patients with suicidal ideations that finding was even more pronounced (86%) with the highest frequency of completely not visible TCS raphe finding (72%). Data showed that altered echogenicity of the BR is frequent in patients with suicidal ideation.

Fifty-five adults with CP, in levels I to V of the Gross Motor PF-562271 supplier Function Classification System (GMFCS) (31 men and 24 women; mean age, 37.5±13.3y; range, 18–65y), were recruited from the Central Remedial Clinic, a national center for the treatment and care of people with disabilities, and through general practitioners nationwide. The GMFCS is a 5-point scale that distinguishes between levels (I–V) of motor function on the basis of functional mobility and the need for assistive technology, particularly mobility aids.16 Adults with a severe intellectual disability (intelligence quotient <35) and pregnant women were excluded from participating in this study. The register of the center

was searched for eligible persons, resulting in study invitations being sent to 263 adults with CP. Forty-three adults with CP responded and consented to participate. Information about the study and a request to recruit eligible persons was sent to 1367 general Selleck MS 275 practitioners; 7 participants were recruited by this method. The remaining participants were recruited by word of mouth. Ethical approval for this study was granted by the Faculty of Health Sciences’ ethics committee and the Central Remedial Clinic’s ethics committee. All participants, and their guardians

in the case of adults with mild-to-moderate intellectual disability, provided written informed consent before data collection. Anthropometric measures including stature, body mass, WC, and hip circumference were obtained. In the case of nonambulatory participants, stature was predicted from knee height.17 Knee height was measured with the knee and ankle held at 90°, from the posterior surface of the thigh, just proximal to the patella, to the sole of the foot, using calipers. WC was measured, on bare

skin, to the nearest 0.1cm midway between the lower rib margin and the iliac crest at the end of gentle expiration. Hip circumference Tacrolimus (FK506) was measured to the nearest 0.1cm at the end of gentle expiration around the maximum circumference of the buttocks. WC and hip circumference were measured on ambulatory participants in standing and on nonambulatory participants in supine lying positions. The mean of 2 measurements was calculated for both WC and hip circumference. Overweight and obesity were defined as BMI ≥25kg/m2 and ≥30kg/m2, respectively. Central obesity was defined as WC ≥80cm and ≥94cm for women and men, respectively.18 Blood pressure was measured from the right arm or the least affected side, in the case of significant asymmetry, using the Omron 705 IT blood pressure monitor.a The Omron 705 IT has demonstrated excellent validity in adults.19 Participants rested in a seated position with their backs supported for at least 5 minutes before 3 measurements were taken at 1- to 2-minute intervals. The average of the last 2 measurements was used in data analysis.

2), using the proportion calculated by MONERIS, which was vice versa used to estimate the historical river loads. MONERIS allows simulation and tracking of nutrients from the emission source through the environment to the river mouth. It is based on a geographical information system (GIS), which includes various digital maps and extensive statistical information. MONERIS is applied to calculate riverine nutrient emissions from the German Baltic river

basin, considering also nutrient retention in the river and providing monthly loads at the river mouth. Behrendt and Dannowski [3] and Venohr et al. [53] present details about the model. A comparison between observed and model simulated N and P loads for the period 1983–2005 is documented in Venohr et al. [52]. MONERIS model simulations SB203580 in vivo for the years around 1880 were based on historical statistic data sets and compiled literature data. The German Baltic river basins cover an area of 28,600 km2 or about 2% of the Baltic Sea catchment [23]. In 1880, arable land covered 55%, forests 18% and grassland 15% of the catchment. Agriculture Tacrolimus mouse already covered an area comparable to the present situation, but was still not intensified with only limited application of manure. The nitrogen surplus (difference between

fertilizer application and removal with harvest) was still close to zero. Tile drainage and sewer systems were already in Teicoplanin place. The total human population in the catchment was 1.4 million, roughly 50% less than today. Details about approach and results are described in [27]. Two ERGOM-MOM model simulations were carried out. The first covered the present situation between 1970 and 2008. The average annual German Baltic riverine loads, for example, for the years 2000 until 2008 were about 21,100 t total nitrogen (TN) and 474 t total phosphorous (TP) with an N to P relationship of 39. The second simulation covered the historical situation, using the loads provided by MONERIS for the years around 1880. The historic annual German Baltic riverine loads were 5127 t TN and 227 t

TP (molar N/P=44). The historic run covered the years 1875 until 1885. In subsequent calculations, the simulation results were averaged over the period 2000 until 2008 resp. 1881 until 1885 to reduce the effects of interannual variability and the model dependency on initial starting conditions. To calculate maximum allowable German nutrient inputs and subsequent target concentrations for German rivers, a simplified, spatially integrated approach was used, that allows a direct comparison to existing MAI and the BSAP. The annual DIN and DIP loads and average chl.a concentrations were extracted from model simulations for an area, which is known to influence water quality in the German Baltic Sea (9.5°–14.8°east, 53.6°–55.35°north). To extend the data set, earlier ERGOM-MOM simulations [20] and [31] were additionally considered. Chl.