Treatment of animals with Pyl A

alone increased NF-κB activity in the myometrium, which was enhanced with co-administration of LPS (Fig. 6a). The inability Selleckchem Osimertinib of Pyl A to inhibit NF-κB implies that CRTH2 is not involved in the mechanism of 15dPGJ2-mediated inhibition. In support of this, we demonstrated that CRTH2 is not required for 15dPGJ2-mediated inhibition of NF-κB in human amniocytes, myocytes and lymphocytes.[41] Surprisingly, myometrial COX-2 protein levels remained unchanged 4·5 hr post treatment in all groups. As preterm labour was typically induced following LPS/Pyl A treatment at 5·8 hr (SEM ± 0·7) it was expected that any COX-2 up-regulation in the myometrium should have already been apparent by 4·5 hr post treatment. It is possible that COX-2 was already up-regulated before intrauterine injection in preparation for term labour, which is one limitation of using a model at E16. Progesterone withdrawal in the mouse occurs late E16 and so downstream activation of pro-labour genes is not likely to have been initiated in our model.[44] Consistent

with this the majority of labour-associated Midostaurin ic50 proteins such as PGE2, PGF2α, the oxytocin receptor and Connexin-43 are not significantly up-regulated until E18.[45, 46] We have shown, however, that COX-2 is suppressed in pregnancy and is up-regulated from E16, which was not increased further in term labour.[47] We further explored the possibility that, despite seeing no change at the protein level, COX-2 was still activated by LPS and LPS plus Pyl A. Messenger RNA was indeed increased

in LPS-treated mice, and was further increased with co-injection of Pyl A (Fig. 6e). COX-2 requires peroxidases for activation and the endogenous peroxide tone of smooth muscle cells can be mimicked by nitration.[48] Previous studies have shown that peroxynitrite increases the activity of COX-2 with no alteration of COX-2 protein expression.[49, Resveratrol 50] Consistent with our results, Aisemberg et al.[51] demonstrated an increase in LPS-induced mRNA COX-2 with no effect at the protein level. It is plausible that this is a result of LPS-induced NO leading to the formation of peroxynitrite, which in turn, activates COX-2 without alteration of protein expression. Alternatively, it is also plausible that the nitrated form of COX-2 is not recognized by the COX-2 antibody. Analysis of pup brain extracts collected from LPS-treated dams revealed a decrease in levels of phosphorylated p65 (ser 536). It is thought that this may reflect protein degradation induced by the pre-terminal state of the live pups (Fig. 6b). A significant increase in in utero fetal viability was achieved with Pyl A treatment (Fig. 5a) but this was not associated with altered NF-κB activity. This also highlights the contrasting effects of Pyl A compared with the 15dPGJ2 because we have previously shown that 15dPGJ2 inhibits NF-κB in the pup brain of dams treated with LPS.

We observed no changes in lymphocyte motility or diapedesis (Fig. 5A). Analysis of live-cell videomicroscopy indicated a similar fraction of lymphocytes encountered at least one interendothelial junction during movement on control or ND-treated monolayers, (83±5% versus 87±3% (mean±SEM);

p=NS, n=5 independent experiments). Further, analysis of immunofluorescence images of co-cultures of lymphocytes adherent to EC monolayers, fixed after 10 min of applied shear, was consistent with the videomicroscopy results. DMXAA manufacturer We observed no difference in the fraction of adherent lymphocytes in contact with VE-cadherin stained junctions between control and ND-treated monolayers (76±4% versus 75±5% (mean±SEM); p=NS, n=6 independent experiments).

These results indicate that loss of cortical endothelial MT does not influence movement of lymphocytes to the interendothelial junction, suggesting that endothelial MT play a role in lymphocyte interpenetration of adjacent EC. The location of lymphocytes within the interendothelial junction, in EC treated with ND or vehicle reagent, was analyzed by confocal microscopy as described in Fig. 3 legend. Data from lymphocytes adherent to control (n=367) or ND-treated (n=341) monolayers in three independent experiments was pooled. Analysis of the position of the lymphocytes revealed that the fraction of lymphocytes in a suprajunction position was 1.3-fold higher among MT-depolymerized EC monolayers versus control (Fig. 5B; p<0.01). The fraction that completed diapedesis in the ND-treated group selleck chemical was reduced to ∼60% of the DMSO-treated group (Fig. 5B; p<0.01). Thus, both videomicroscopy and confocal imaging techniques indicate that

endothelial MT are required for efficient diapedesis, but are not essential for lymphocyte locomotion on the EC surface. Further, loss of IQGAP1 expression and MT depolymerization both cause lymphocytes to accumulate above the AJ. Leukocyte diapedesis is associated with specific and transient gap formation in AJ 13, 14, 18; hence, we investigated whether loss of EC IQGAP1 or MT depolymerization affected gap formation associated with suprajunction-localized lymphocytes. We observed 22±3% of lymphocytes adherent to control monolayers were associated with find more a gap >2 μm in diameter. Neither IQGAP1 knockdown nor ND treatment change the fraction of lymphocytes associated with VE-cadherin gap formation (110±36% versus 98±15% of control (mean±SEM); siIQGAP1 versus ND treatment; four independent experiments). Further, we examined the frequency of gaps enriched in PECAM-1 distributed around transmigrating lymphocytes. In these experiments, we studied TEM of PECAM-1−/dim memory T cells. We observed 32±9% ((mean±SEM); three independent experiments) of lymphocytes migrating across control EC monolayers were associated with a VE-cadherin gap enriched in CD31 (Supporting Information Fig. 6).

Indeed, as shown in Fig. 6B similar Foxp3 staining could be observed

in the lamina propria of TNBS-treated mice that received PI when compared to TNBS-treated mice that received saline suggesting that Foxp3+ T cells are normally present in the lamina propria of PI-treated mice. Our data demonstrate that the phospholipid PI inhibits T-cell proliferation and BGB324 molecular weight differentiation but does not affect Treg differentiation. In vitro experiments established that PI strongly inhibited PMA-stimulated T-cell activation. Moreover, T cells stimulated with plate-bound anti-CD3 and soluble anti-CD28 antibodies were also effectively inhibited by PI. The latter experiments excluded that PI acted through interference with processes such as transmembrane diffusion of PMA. Inhibition was dose dependent and did not involve anergy, apoptosis or deletion as reflected by the fact that suppression selleck compound was reversible when PI was removed (Supporting Information Fig. 3). Moreover, suppression was not restricted to a specific subset of T cells as both CD4+ and CD8+ T-cell activation was inhibited. In contrast, PI did not affect T-cell proliferation indirectly by inhibiting antigen-presenting DCs, as loading of DCs in the presence of PI before co-incubation did not

inhibit T-cell activation. These data strongly implicate an effect of PI on the intracellular pathways that are associated with T-cell activation. By determining phosphorylation activity of various intracellular checkpoints of T-cell activation we established that PI exerted its effects on the PKC–MAPK pathway. As we found decreased ERK1/2, P38 and JNK phosphorylation upon PI treatment, we hypothesize that PI targets Pyruvate dehydrogenase one of the common inositol lipid pathways that precedes these downstream routes. Recently, it was established that inositol lipid signaling is regulated through cellular expression of a class of PI-transfer proteins PI-TPα and PI-TPβ 11. Modulation of this signaling has been associated with changes in cellular proliferation. In particular, overexpression of PI-TPα in fibroblasts induces an increased growth rate, whereas the growth rate of PI-TPβ overexpressing cells

is decreased 12, 13. Future studies into the specific regulation of expression of such transfer proteins may lead to the development of novel PI-based immunosuppressants. The inhibition of signal transduction in T cells cultured with PI led to reduced IL-2 mRNA expression and low levels of IL-2 protein release, which abrogated T-cell proliferation. In consequence, PI inhibited inflammatory CD4+ Th cell proliferation and cytokine release. Crucially, Foxp3+ Treg differentiation was not aborted by addition of PI, which is a pivotal characteristic for a potential immunosuppressant. As such, the immunosuppressive effects of PI share more similarity with the inhibitor rapamycin, which preserves Foxp3+ T cells while inhibiting inflammatory responses than with cyclosporine, which suppresses both inflammatory and Tregs 14, 15.

Because familiarity preferences like this emerge when infants are relatively slow to process a habituation stimulus, the data support the interpretation that mental rotation of dynamic three-dimensional stimuli is relatively difficult—but possible—for 3-month-old males. Interpretation of the sex differences observed in 3- and 5-month-olds’ performances is discussed. “
“Past studies have identified individual differences in infant visual attention based upon peak look duration during initial exposure to a stimulus. Colombo and colleagues found that infants that demonstrate brief

visual fixations (i.e., short lookers) during familiarization are more likely to demonstrate evidence of recognition memory during subsequent DAPT purchase stimulus exposure than infants that demonstrate long visual fixations Erlotinib (i.e., long lookers). This study utilized event-related potentials (ERPs) to examine possible neural mechanisms associated with individual differences in visual attention and recognition memory for 6- and 7.5-month-old infants. Short- and long-looking

infants viewed images of familiar and novel objects during ERP testing. There was a stimulus type by looker type interaction at temporal and frontal electrodes on the late slow wave (LSW). Short lookers demonstrated an LSW that was significantly greater in amplitude in response to novel stimulus presentations. No significant differences in LSW amplitude were found based on stimulus type for long lookers.

These results indicate deeper processing and recognition memory of the familiar stimulus for short lookers. “
“Despite the use of visual habituation over the past half century, relatively little is known about its underlying processes. We analyzed heart rate (HR) taken simultaneous with looking during infant-controlled habituation sessions collected longitudinally at 4, 6, and 8 months of age with the goal of examining how HR and HR-defined phases of attention change across habituation. There were four major findings. First, the depth and topography of decelerations and proportion of sustained attention (SA) enough did not vary across habituation at any age, which suggested (in contrast to the tenets of comparator theory) the persistence of substantial cognitive activity at the end of visual habituation. Second, attention termination (AT) robustly declined across trials, suggesting that, contrary to prior thinking, AT might be a sensitive indicant of visual learning. Third, infants at all ages showed an HR increase (startle) to stimulus onset on the first trial, the magnitude of which was associated with subsequent delayed HR deceleration and less SA; thus, stimulus events affect processing during trials. Finally, mean overall HR reliably increased across trials for all ages. This last finding implies the need to distinguish between “phasic” HR changes (e.g.

9B). Of note, the level of KLRG1 expression by NK cells from KLRG1 TG mice was considerably higher when compared with NK cells from WT mice (MF 186 versus 43)

(Fig. 9C). Metformin These data indicate that high KLRG1 expression levels by NK cells are required for E-cadherin-mediated inhibition in the murine system. It is noteworthy that functional activity of human KLRG1+ NK cells could be significantly inhibited by E-cadherin in the same assay system used here with K562 target cells 24. Since natural KLRG1 expression by ex vivo isolated NK cells from humans and mice are similar, these data point to a difference in the inhibitory capacity of mouse and human KLRG1. In an attempt to unravel the role of KLRG1 in vivo, we generated and characterized KLRG1-deficient mice. Although a number of different infection models and assays systems were used, we failed to observe an effect of KLRG1 deficiency on NK and T-cell differentiation and function in vivo. How can these “negative” findings be rationalized? In the targeting vector used for homologous recombination, the Klrg1 gene was disrupted by insertion CH5424802 manufacturer of a LacZ/neomycin expression cassette into the third exon. Appropriate homologous recombination was confirmed by Southern blotting and

lack of KLRG1 expression was also verified at the mRNA and at the protein level. Alternatively spliced transcripts of the Klrg1 gene are detectable at a low level but none of these transcripts is predicted to give rise to a protein with residual KLRG1 activity since they either lack a transmembrane region or lead to a frame shift in the extracellular part 2. Thus, the lack of a phenotype in KLRG1 KO mice is very unlikely due to imperfect ablation of the Klrg1 gene. Of note, KLRG1 is present in the genome as a single copy gene 6

and closely related receptors are not known. PLEKHM2 Inhibition of T and NK-cell function by antibody- or E-cadherin-mediated ligation of KLRG1 has been documented by several groups 21–23, 25, 26. It is, however, important to stress that all inhibition experiments published so far in the murine system have been performed with retrovirally transduced cell lines or transgenic lymphocytes that over-express KLRG1. We demonstrate here that E-cadherin expressed by K562 target cells could only inhibit NK cells from transgenic mice over-expressing KLRG1 but not from normal mice. This indicates that the inhibitory potential of mouse KLRG1 is rather weak and requires high levels of expression. It is therefore possible that the weak inhibitory signal through KLRG1 was overruled by strong activation stimuli in the infections models used here. Model systems that are accompanied with lower activation of immune cells may therefore be suited better to unravel the function of KLRG1 in vivo.

The effect of smoking on the immune response and thereby the kynurenine pathway is multi-faceted, and may reflect the opposing nature of cigarette smoking as a proinflammatory factor and the immunosuppression mediated by nicotine [25]. This is the largest Z-VAD-FMK solubility dmso community-based study investigating biological

and lifestyle determinants of plasma levels of neopterin, KTR and kynurenines. The large sample size and comprehensive data on a large panel of kynurenines and lifestyle factors are unique. The observed plasma concentrations were similar to those reported in another large cohort study [41]. In addition to self-reported smoking behaviour, plasma cotinine provided reliable information on recent nicotine exposure. The cohort enabled us to compare levels of kynurenines and related markers of inflammation between two distinct age groups (46–47 and 70–72 years). However, we could not evaluate the effect of age as a continuous variable, or in other GSK-3 phosphorylation age groups. Lastly, the associations with physical activity might be attenuated, as physical activity was not assessed using a validated physical activity questionnaire. Nevertheless, to the extent of our knowledge, this is the first study that addresses habitual physical activity

as a determinant of plasma neopterin, KTR and kynurenines. Neopterin and KTR are both markers of cellular immune activation, whereas some kynurenines have immune modulatory effects. We observed strong

positive associations between these markers and metabolites with age and renal function, indicating that neopterin, KTR and the kynurenines are sufficiently responsive indices to capture the low-grade inflammation that occurs in the elderly. Additionally, KTR and most kynurenines were higher in overweight/obesity, and several kynurenines were associated inversely with smoking. The data also demonstrate that KTR GNAT2 and most kynurenines may reflect the low-grade inflammation present in obese subjects, whereas the inverse association between several kynurenines and smoking potentially reflects the complex effect of smoking in immune functions. Such knowledge highlights potential confounding in epidemiological and clinical studies, but also motivates the inclusion of markers of cellular immunity to disentangle various components of systemic inflammation in the pathogenesis of chronic diseases such as cardiovascular disease and cancer. This work was supported by the Norwegian Research Council (project number 204650), and funded partly by the non-profit ‘Foundation to Promote Research into Functional Vitamin B12 Deficiency’. We thank Marit Krokeide, Anne-Kirstin Thoresen and Gry Kvalheim for their technical assistance. The authors declare that there are no conflicts of interest. Table S1.

Tbet was expressed at a significantly higher level in the colons from the Aire-group (Fig. 4B). No differences were found in the expression of other T helper cell (Th) cell lineage genes GATA3 and

RORγT. Finally, as a systemic marker of ongoing inflammation and colitis [40] we measured the concentration of acute Silmitasertib ic50 phase protein serum amyloid protein (SAP) in the recipient mice. Compared with both Aire−/− and Aire+/+ control animals without cell transfers, both groups of recipients had elevated plasma levels of SAP, but there was no statistically significant difference between the groups (Fig. 4C). The surprising lack of clinical disease, despite autoantibodies and other signs of autoreactivity in the Aire-group, prompted us to look at Tregs in the recipients. One month after the cell transfer, the proportion of circulating Foxp3+ cells among all CD4+ cells was comparable in both groups (control-group 6.2 ± 2.0% and Aire-group 4.7 ± 0.9%, difference not significant). At the time of termination, the frequency of circulating Foxp3+ cells remained similar in both recipient groups (Fig. 5A). However, the frequency of MLN8237 research buy circulating Foxp3+ cells expressing the cell cycle marker Ki-67 was significantly higher in the Aire group (Fig. 5B). To test whether this higher rate of proliferation resulted in increased accumulation of Treg cells

in the Aire group we then analysed the frequency of Foxp3+ cells in the recipients’ lymphoid tissues. In spleen, the frequency was similar in both groups (16.6 ± 4.1% and 17.5 ± 6.1% in the control and Aire group, respectively). In the mesenteric lymph nodes, in contrast, the frequency of both Foxp3+ cells, and the fraction of Treg

cells expressing Ki-67, was much higher in the Aire group (Fig. 5C,D). Moreover, the amount of Foxp3 mRNA in the colon tissue, normalized against TCR Cα mRNA, was higher in the Aire group recipients (Fig. 5E). Together, these data indicate that Treg cells hyperproliferated in the Aire group recipients, Calpain accumulating in higher numbers to potential sites of inflammation. The importance of Aire to the development of central tolerance is clearly established [17, 20], but there is also increasing evidence that Aire is needed for maintaining peripheral tolerance [23, 24, 41]. Our model of LIP allowed us to determine how much of the Aire−/− phenotype is duplicated, when T cells that have matured in the absence of Aire are exposed to autoimmunity-provoking signals within an Aire-sufficient peripheral environment. Adoptive cell transfers have previously been carried out both using bulk lymphocytes and selected subsets of T cells. In our experiments, we chose to do the former. In several murine models of autoimmunity, such bulk transfers to lymphopenic recipients have been reported to successfully transfer the disease [28, 42–44], and in some models, the co-transfer of B and T cells are indeed required to trigger autoimmunity [45].

Using DNA-cytometric analysis, Ihrler et al.’s [37] study described the presence of chromosomal alterations in salivary gland MALT lymphoma in SS. Regarding the key role of BAFF in SS proposed by some authors [4,39], the assessment of BAFF levels in serum is an exciting field for future research. Our study showed a high prevalence (86·7%) of B cell clonality in patients with SS and a direct relationship with the degree of focal lymphocytic infiltrates. In healthy control groups, we observed a direct correlation between CHIR-99021 solubility dmso the degree of CS and the presence of oligo- or monoclonal bands. Therefore, this study supported the hypothesis that an increasing

number of patients with different degrees of CS may result in clonal B infiltration of the gland, showing an association between the severity of the MSG inflammation pattern and the presence of clonality. The finding of clonality in samples from this group of individuals is interesting, and possible explanations of these results are: (i) the development of reactive clonal population, distributed widely in the salivary glands, as has been reported in other studies [33,34]; and (ii) PCR is a very sensitive Mitomycin C mouse technique, and could detect a few cells among a normal cellular background. According to our results, we show in this paper that the detection of B cell clonality by

PCR in MSG of SS patients is a predictor of clonal expansion. Clonal expansion during chronic gland inflammation of B cell mutations takes place regularly, accompanied by mutations of tumour-suppressor genes, p53 mutations and a high level of BAFF expression. Together, these alterations constitute a risk factor for the development of lymphoma in SS patients [4–6,29,30,34,40]. We conclude that the presence of B cell clonality in MSG can be used as an index of an altered microenvironment, which could enable the development of lymphoma in SS patients. This research was supported by funds of the Public Institute of Health from Chile, Bagó Laboratory and Chile Laboratory. All authors declare

selleck no conflicts of interest. “
“It is now well established that allergic diseases have an extremely high prevalence in developed societies, and are increasing in emerging countries. In fact, allergy is probably the most prevalent immunological disease. It is currently estimated that up to 30% of Europeans suffer from allergic rhinitis or conjunctivitis, while up to 20% suffer from asthma and 15% from allergic skin conditions 1. The worldwide numbers are equally worrying. Almost half a billion suffer from rhinitis 2, 3 and approximately 300 million from asthma 4. Compared with other chronic diseases, allergic diseases are more common than Parkinson’s, Alzheimer’s, stroke, coronary heart disease, cancer or diabetes.

39 α-lipoic ABT-263 cell line acid (α-LA) is also found naturally in mitochondria and acts as a critical coenzyme for the mitochondria enzymes pyruvate dehydrogenase and α—ketoglutarate dehydrogenase.40 Its reduced form, dihydrolipoic acid (DHLA), and other metabolites have strong antioxidant effects as ROS scavengers and act as chelators of transition metals.41 In a PBOO study, CoQ10 plus α-LA treatment significantly

increased bladder contractility in vitro, decreased bladder wall protein carbonylation and nitration, increased mitochondrial function, prevented intramural nerve degeneration and diminished detrusor smooth muscle hypertrophy in rabbit bladder.26 Specifically, bladder voiding contraction can be separated into two phases: an initial peak phase and a second tonic phase.42 The initial peak response CYC202 was supported energetically by extant cellular ATP stores, whereas the tonic phase required active mitochondrial oxidation of substrates to generate energy. Ability of the bladder to sustain contraction is directly related to the availability of energy produced by mitochondrial electron transport and oxidative phosphorylation.42 Decompensation of bladder from extended obstruction may

be mediated by breakdown of mitochondrial function. Both CoQ10 and α-LA are essential mitochondrial components in respiratory chain and pyruvate-dehydrogenase complex. Combination therapy may target several common pathways in mitochondrial dysfunctions. Following 4 weeks PBOO, both choline acetyl-transferase activity (an indicator of cholinergic function) and neurofilament amounts decreased significantly and diminished further after 7 weeks of obstruction. PBOO also significantly increased both protein nitration and carbonylation following obstruction, especially after 7 weeks obstruction. CoQ10 and α-LA are both strong antioxidant reagents in nature. Treatment with CoQ10 plus α-LA significantly attenuated protein carbonylation Tangeritin and nitration, indicating that these medications

may work through an antioxidative effect. The antioxidative function of CoQ10 is likely to occur by providing hydrogen equivalents to reduce peroxyl and/or generation of alkoxylradicals.39α-LA has also been proven to reduce lipid peroxidation by enhancing the activity of glutathione peroxidase and SOD, which improves the efficiency of the endogenous antioxidant systems.43 A combination of these two strong antioxidants thus prevents free radical induced tissue damages subjected to PBOO. Ischemia/reperfusion injury is also involved in bladder overdistention injury. It has been known for a long time that bladder overdistention reduces blood perfusion of the bladder.44 Blood flow is resumed following emptying and decompressing the urinary bladder. Reperfusion of the overdistended and ischemic urinary bladder might induce reperfusion injury. In a rabbit overdistention model, Lin et al.

Gp96, a 96-kDa glycoprotein, is a member of the HSP90 family and resident in the endoplasmic reticulum Omipalisib datasheet (ER) [12]. It possesses a signal peptide of 21 amino acids at the N-terminal region of the protein which is cleaved cotranslationally, while the C-terminal contains KDEL, an ER-retention sequence [13]. Gp96-specific interaction with CD91 receptor which expressed on professional APCs mediates endocytosis [14]. Receptor-mediated endocytosis of gp96 molecule leads to MHC class I-restricted re-presentation of gp96-associated peptides [15]. Several studies have established the ability of gp96 to activate innate immune responses and thereby influence the

outcome of adaptive immune responses. Gp96 is able to mediate maturation of DCs in a TLR4-dependent manner, as determined by upregulation SP600125 cell line of MHC class II, CD86 and CD83 molecules, secretion of pro-inflammatory cytokines IL-12 and TNF-α and enhanced T cell stimulatory capacity. The interaction of gp96 with DCs leads to the preferential expansion of antigen-specific CD8-positive

T cells in vitro and in vivo [16, 17]. It was demonstrated that amino acid sequence 1–355 of gp96 is sufficient to bind peptides and mediates the uptake of peptides into the endosomal compartment of APCs. In comparison with the full-length gp96, the N-terminal fragment up-regulates the same costimulatory receptors and induces secretion of the same cytokines [18, 19]. Furthermore, co-administration of N-terminal fragment of gp96 along with Hepatitis-B surface antigen (HBsAg) enhances the humoral immunity induced by Y-27632 2HCl HBsAg [20] and CTL immune responses to Hepatitis-B-Virus (HBV) peptide [21]. Further study indicated the construction

of highly immunogenic fusions by linking the N-terminal fragment of gp96 to HBV antigens [22]. Altogether, these data imply that the N-terminal fragment of gp96 performs the same adjuvant activity to enhance the potency of vaccines as the full-length gp96. Indeed, the studies in animal model revealed that DNA [23] or protein [24, 25] vaccination with full-length antigen co-linked to different HSPs elicit antigen-specific immune responses. In the current study, the humoral and cellular immune responses as well as the protective anti-tumour immunity using the adjuvant-free recombinant (r) HPV16 E7-NT-gp96 fusion protein were evaluated and compared to rE7 alone in tumour mice model. Mice and cell line.  Female C57BL/6 mice, 6–8-weeks old, were obtained from breeding stock maintained at the Pasteur Institute of Iran. TC-1 (ATCC number: CRL-2785) tumour cell line was prepared from primary lung epithelial cells by co-transformation with HPV16 E6, HPV16 E7 and ras oncogenes [26]. The TC-1 cancerous cell line was cultured in RPMI 1640 (Sigma, St.