A detailed phenotypic characterization of induced CD8+Foxp3+ T cells revealed high expression of classical Treg markers including CD25, GITR and CTLA4, consistent with previous reports 17, 31 and likely reflecting T-cell activation, although one study reported low CD25 expression on CD8+Foxp3+ T cells 38. Interestingly, the classical

Treg markers CD73 and CD103 were selectively expressed by induced CD8+Foxp3+ T cells, underlining that their expression is dependent on TGF-β, RA and/or Foxp3. In line with this, CD8+ T cells deficient in TGF-β signaling fail to up-regulate CD103 in a GVHD model 39, and Foxp3 has been shown to directly bind the CD103 promoter 40. However, Foxp3-independent mechanisms can also activate CD103 3, consistent with the only mildly reduced induction of CD103 expression in stimulated T cells Pictilisib nmr from DEREG×Rag1−/−×OTI×Sf mice (Supporting Information Fig. 3C). CD8+Foxp3+ T cells only displayed little suppressive capacity compared with CD4+Foxp3+ Tregs, and CD8+Foxp3− T cells showed similarly low suppressive activity in vitro. Furthermore, adoptive transfer AZD0530 of induced CD8+Foxp3+ T cells did not ameliorate disease in an OVA-based allergic airway inflammation model (data not shown). Previous studies have reported the suppressive capacity of TGF-β-induced

CD8+ T cells 17, 31, 34, 38, which in principle does not contradict our data. First, several studies did not compare the strength of suppression to that of CD4+ Tregs 31, 34, 38, which depend on Foxp3 3. Second, suppressive CD8+ T cells were isolated either based on CD25 expression 17 (also broadly up-regulated on activated Foxp3− T cells, at least in the absence of IL-6), or were tested without second further separation for suppressive function 31, 38, thereby not allowing for discrimination between Foxp3+ and Foxp3− subsets. Third, DC or agonistic αCD28

antibodies were used during in vitro differentiation in all these studies. Therefore, it cannot be formally excluded that the low suppressive function observed in our study is caused by the lack of signals provided by either DC or αCD28. However, this would underlie Foxp3-independent mechanisms, since CD8+Foxp3+ T cells can be efficiently generated without co-stimulation (Fig. 1). Strikingly, co-stimulation even represses Foxp3 induction in CD8+ T cells (Fig. 2A and B) suggesting that CD80/CD86–αCD28 would rather modulate suppressive activity in a Foxp3− subset. In sum, our results suggest that Foxp3 alone is not sufficient to confer strong suppressive activity to CD8+ T cells. Although transgenic mice with forced overexpression of Foxp3, but not WT mice, were described to harbor suppressive CD8+ T cells, Foxp3 was similarly considered as implicated but not sufficient to confer suppressive activity in a previous study 41.

This “outside-in” signaling pathway requires ITAM signals from DAP12 and FcRγ, and also involves early effectors such as the Src family kinases and Syk in neutrophils and macrophages [14, 15]. Because β2 integrins signal through

ITAM adapters in myeloid cells, we hypothesized that β2 integrin signaling may also inhibit TLR responses. There have been conflicting reports in the literature regarding the influence of β2 integrin signaling on TLRs, with some studies demonstrating that β2 integrins can promote TLR-induced inflammation [16-18], whereas others have reported negative roles for these integrins in TLR responses [19, 20]. Therefore, the nature in which β2 integrins interface with TLR activation and cytokine secretion is complex PD-1/PD-L1 targets and unclear.

To better define the contribution of β2 integrins to regulation of TLR signaling, we have examined inflammatory responses in the absence of all β2 integrins. Here we demonstrate that deletion of all β2 integrins rendered myeloid cells hypersensitive to TLR stimulation in vitro and in vivo, showing an inhibitory role for β2 integrins in TLR responses. Furthermore, Sunitinib mw we examined potential direct and indirect mechanisms by which β2 integrins caused this inhibition, and found that β2 integrins have a direct effect on IκBα degradation that was pronounced in β2 integrin-deficient cells through both early and late phases of TLR stimulation, thus implicating β2 integrin signals in inhibiting NF-κB pathway activation to calibrate inflammatory responses. The four β2 integrins, LFA-1 (lymphocyte function-associated antigen 1, αLβ2), Mac-1 (macrophage-1 antigen, αMβ2), CR4 (αXβ2), and CD11d-CD18 (αDβ2) are heterodimers that consist of distinct CD11 alpha subunits in association with the common

beta chain, CD18 (β2), which is encoded by the Itgb2 gene [21]. To examine whether β2 integrin signaling regulates TLR responses, we compared the cytokine secretion profiles of bone marrow-derived (BM-derived) macrophages from wild-type Niclosamide (WT) and Itgb2−/− mice, which are deficient in CD18 and thus are unable to express any of the β2 integrins on the cell surface (Supporting Information Fig. 1A) [22]. Despite the inability of Itgb2−/− BM-derived macrophages to express Mac-1, these cells exhibited surface F4/80 expression and upregulated MHC II in response to IFN-γ treatment (Supporting Information Fig. 1A and B), demonstrating that they were bona fide macrophages. Furthermore, β2 integrin-deficient macrophages exhibited similar or slightly lower levels of cell surface TLR2, TLR4, and Dectin-1 protein and TLR9 mRNA (Supporting Information Fig. 1C and D). To determine how β2 integrin signals influence TLR activity, we stimulated Itgb2−/− BM-derived macrophages with a panel of TLR agonists, including LPS (TLR4), CpG B DNA (TLR9), and zymosan (TLR2).

The organization of these gVLR genes differs depending on the gene and species (2b). The possible combinations of VLRA and VLRB are estimated to generate a potential repertoire that is almost equivalent to the TCRs

and BCRs of jawed vertebrates, (> 1014) [22]. This observation suggests that VLRs are the antigen receptors of jawless vertebrates. Consistent with this, lampreys immunized with human erythrocytes or anthrax spores of Bacillus anthracis produce antigen-specific soluble VLRB molecules that act as antibodies [22], [23]. These observations indicate that, despite their lack of structural similarity to the selleck screening library antigen receptors of jawed vertebrates, VLRs function as antigen receptors in jawless vertebrates. During development of LLCs, LRR modules are inserted into the gVLR gene by a gene conversion-like

mechanism (2c) [19], [24]. Multiple LRRNT-, LRR1-, LRRV-, LRRVe-, CP- and LRRCT-encoding modules are located proximally to the gVLR gene. A homologous sequence is used to prime the insertion of those modules during VLR assembly. The sequences located at the ends of the most newly copied LRR module determine the next LRR module. In this way, LRR modules are unidirectionally inserted into the gVLR gene. Although, monoallelic assembly of the VLRA and VLRB genes occurs in the majority of cases, diallelic assembly has BAY 80-6946 been observed in a few cases [25]. In such instances, one mature VLR gene encodes a functional VLR structure, while the other does not. The

unsuccessful mature VLR gene contains an in-frame stop codon. These observations indicate that an inhibitory feedback mechanism regulates VLR assembly. The molecular mechanism of VLR assembly is still unknown. However, two CDAs, CDA1 and CDA2, have been identified as candidate molecules that may mediate gene conversion [19]. Generation of antibody diversity by gene conversion in birds, rabbits and cattle requires AID, which belongs to the apolipoprotein B mRNA editing enzyme, catalytic PRKACG polypeptide family of molecules [26]. Phylogenetic analysis and secondary structure prediction suggest that AID and CDA1 are more closely related to each other than are AID and CDA2. Over-expression of the CDA1 molecule in yeast confers a mutagenic phenotype and increases the rate of intragenic recombination. Previous reports have revealed that CDA1 and CDA2 are expressed in VLRA+ and VLRB+ LLCs, respectively [27]. Thus, the jawless vertebrate CDA1 and CDA2 molecules may control gene conversion-like processes in VLRA+ and VLRB+ LLCs, respectively. Jawless vertebrates possess both soluble and membrane-bound forms of VLRB [17], [28]. Soluble VLRB antibodies are organized into pentamers or tetramers of dimers, similarly to immunoglobulin M of jawed vertebrates. The cysteine residues that are located in the 3′-invariant stalk region are required for VLRB antibodies to form oligomers.

Screening based on title and abstract identified 56 citations for full-text review (Fig. 1). Additional five studies[25-27, 39, 53] were identified from reference lists of the identified articles and from other databases. Of the 56 potentially relevant articles,

32 were excluded for reasons given in Figure 1, leaving a total of 24 studies[24-47] that met the inclusion criteria. Twenty one studies[24-30, 32, 34-38, 40-47] reported associations Ku-0059436 chemical structure between use of statins and AKI, and 14 studies[28, 31-35, 37, 39-41, 43-46] reported associations between use of statins and AKI requiring RRT. Five studies[24-28] used RCT design, and the rest applied a cohort design.[29-47] Only one RCT[28] defined AKI as the primary endpoint. The other four RCTs defined postoperative thrombocytosis,[24] postoperative inflammatory responses,[25]

postoperative myocardial injury,[26] and the number of postoperative endothelial progenitor cells[27] as primary endpoints. Among the cohort studies, only three used prospective design; the remaining studies were retrospective in design. As for the study population, two studies involved nation-wide populations, while most of the other studies were conducted at one single centre. Among the two population-based studies, one was conducted in Canada,[43] and the other in the USA.[47] We assessed the quality Staurosporine solubility dmso of included studies with the Jadad scale.[54] The study conducted by Prowle JR and colleagues[28]

had the highest score on the Jadad scale. The results were summarized in the Appendix 1 (Table App1). The studies varied in their types of surgery, mean age, and case definition (Table 1). The types of surgery were restricted to cardiac or vascular surgery in most studies. Specific type, dosage, and duration of preoperative statin therapy Adenosine triphosphate were not available in most studies. In contrast to AKI defined by database codes, AKI defined by a pre-specified increase of serum creatinine level was regarded as ‘AKI defined by laboratory criteria’. Among these, there were seven studies[28, 37, 38, 41, 44-46] using AKIN or RIFLE criteria[48, 49] as the definition for AKI. In all studies, the definition of AKI requiring RRT was based on clinical judgment without additional objective laboratory criteria. Specific statin type available i Dosage and duration not available Increase of sCr level > 30% (AKIN stage 1) Atorvastatin 20 mg/day or simvastatin 20 mg/day for at least 6 months Started before surgery Type, dosage and duration not available At least one dose of statin between admission and surgery In the 21 studies reporting the association of statin use and AKI, the incidence of AKI ranged from 1.88%[43] to 52.17%[44] (Table 1). The pooled incidence of AKI for all 21 studies was 4.89%. The pooled incidence of AKI among statin user and nonstatin user were 6.13% and 4.28%, respectively (Table 2).

Moreover, to facilitate the pipeline, during the same period significant infrastructure was emerging in the form of clinical trial networks, within which

clinical studies could be conducted to agreed and standardized designs and protocols. The exemplar of this approach is Type 1 Diabetes TrialNet (http://www.diabetestrialnet.org). There was even significant and demonstrable interest in this disease space being displayed by large pharmaceutical concerns. Consequently, as a result of this constellation of events, in 2007 the clinical trial horizon for type 1 diabetes was viewed with the expectation of success and progress. Some 6 years on, several key questions emerge. What has become of the pipeline and the combination approaches? Using the same format as the 2007 paper, we have updated the data tables with new or contemporary information on trials conducted or in progress Selleckchem Napabucasin at that time, and added information on new and ongoing studies. Information-gathering

is based largely on the US National Institutes of Health-sponsored website ClinicalTrials.gov (http://www.clinicaltrials.gov) and the European equivalent (EU Clinical Trials Register; https://www.clinicaltrialsregister.eu/index.html), as well as our knowledge of the sector. Our analyses include studies conducted in the predisease setting, before diabetes onset, for both antigen-specific and non-antigen-specific approaches [primary (high genetic risk) and secondary (high risk identified by islet cell autoantibody positivity) selleck products prevention studies, Tables 1 and 2, respectively] and trials in which recruitment centres on subjects who have already PAK6 developed disease (intervention studies; Tables 3 and 4, respectively). There is a further

update on trials using combination approaches (Table 5). What have we learned from the clinical trials that have been conducted? Has our general understanding of the disease altered in any respect in the intervening period, such that we might review our therapeutic options? Pre-POINT study: dose finding in children with high genetic risk for type 1 diabetes EudraCT number: 2005-001621-29 Phase II in adults reports preservation of C-peptide at 12–18 months. Phase II in children reports no treatment effect Phase II completed Phase III terminated Anti-CD3 mAb hOKT3g1(Ala-Ala); drug subsequently known as Teplizumab Anti-CD3 mAb ChAglyCD3(TRX4); drug subsequently known as Otelixizumab With the premise that type 1 diabetes is an immune-mediated disorder, most efforts to intervene in disease pathogenesis involve immune-based therapy. Without exception, primary study end-points tend to focus on preservation of β cell function, as measured by stimulated C-peptide production after a standardized food challenge (oral glucose tolerance test, OGTT) or glucagon injection. This is a justifiable criterion that is accepted by regulatory agencies such as the US Food and Drug Administration and European Medicines Agency.

Following stimulation, cells were pelleted, washed, lysed, and immunoprecipitation was performed as described previously [14] using 2.5 μg/mL anti-Lyn or anti-PLCγ2 (Santa Cruz Biotechnology). Samples were run on a FDA approved Drug Library 7.5 or 12% precast SDS-PAGE gel and transferred to a PVDF membrane.

Prior to phosphotyrosine detection, the membrane was blocked and probed with anti-Lyn according to manufacturer’s protocol using a HRP-conjugated light chain specific mouse anti-rabbit IgG (Jackson ImmunoResearch). After the blot was imaged and developed, the membrane was stripped and probed with the anti-phospho-tyrosine antibody described previously. For phospho-PLCγ2 detection, the blot was probed for phospho-tyrosine followed by total protein. To determine the fold increase in phosphorylation for all proteins, the entire protein lane or the protein band was normalized to the total protein. The fold increase in phosphorylation was calculated by multiplying the fold difference in the normalized total protein value by the phosphorylated signal. Fura-Red-AM and Fluo-3-AM ester were purchased from Molecular Probes and dissolved in DMSO as 1 mM and 1.25 mM stock, respectively. Purified B cells were incubated

with 5 μM Fura-Red AM and 2.5 μM Fluo-3-AM buy AZD1208 in PBS containing 5% FCS for 30 min at 37°C in the presence of DMSO control or 10 mM dimedone (dissolved in DMSO). Samples were washed two times with PBS supplemented with 5% FCS and resuspended in the same media containing 10 mM dimedone Teicoplanin or DMSO control. Cells were acquired for 60 s on the FACSCalibur Flow Cytometer and then 10 μg/mL anti-IgM was added to the samples and recording was resumed on the instrument. Endoplasmic reticulum (ER) calcium release and CCE was measured as described by Jia et al. [49]. We thank David Ornelles and Kenneth Grant for their helpful input with the confocal microscopy experiments. This work was supported by NIAID grants RO1-AI068952 and R56-AI073571 to J.M.G and NCI grant R33

CA126659 to L.B.P. K.E.C. was supported by NIAID grant 5T32AI007401-20. The authors declare no financial or commercial conflicts of interest. Disclaimer: Supplementary materials have been peer-reviewed but not copyedited. Figure S1. NAC treatment decreases anti-IgM-induced B-cell proliferation. Figure S2. Dimedone pretreatment decreases cysteine sulfenic acid formation in the total proteome and effector molecules following BCR ligation. Figure S3. NAC treatment initiates ER calcium release and inhibits CCE in B cells. “
“Advanced glycation endproducts (AGEs) of food proteins resulting from the Maillard reaction after cooking or heating may have particular importance in food allergy. The underlying immunological mechanisms are only poorly understood.

32 Urothelium has a basal level of acetylcholine release of non-neuronal origin that increases with bladder distention.35 Further work has linked urothelial acetylcholine to activation of muscarinic receptors and nicotinic receptors with subsequent release of ATP.36 The latter acts on purinergic receptors (P2X) on afferent nerve terminals, possibly providing the important link between acetylcholine and a sensory mechanism selleck inhibitor of action.37 Given this foundation, it seems evident that antimuscarinic medications act during bladder filling and affect sensory activation with little or no effects on motor function

if given at the usual recommended dosages. Higher dosages can produce decreased detrusor contractility and even urinary retention.38 Blockade of muscarinic receptors at detrusor and nondetrusor sites may prevent OAB symptoms and detrusor overactivity without depressing contraction during voiding. selleck chemical Even though the concentration

of antimuscarinic drug is small, it can give some effects on afferent activity. In that case adverse effects also can be decreased Relatively there are few clinical reports about low-dose combination therapy. The evidence level of clinical studies seems low. However, they can hint at a new approach in low-dose combination therapy. For propiverine, 20 mg is thought to be the usual dose and 10 mg to be low dose in East Asia The efficacy and safety of combined therapy with tamsulosin 0.2 mg and low-dose anticholinergic drug (propiverine HCl 10 mg) in BPH patients with OAB symptoms was studied prospectively. One hundred and nineteen

male patients with a prostate volume of 20 mL or greater, IPSS of more than eight, and OAB symptoms were enrolled. Seventy-four patients were treated with tamsulosin 0.2 mg plus propiverine HCl 10 mg (group A) and 45 patients were treated with tamsulosin 0.2 mg only (group B). IPSS, QoL score, voiding volume, Qmax, Urocanase and PVR showed significant improvement after 3 months of treatment. Baseline characteristics between the two groups were not significantly different for any parameter. Changes in the QoL score were statistically significant (−1.9 ± 1.1 and −1.5 ± 0.9 for group A and group B). Changes in all other parameters were not significantly different between the two groups. The authors concluded that combination therapy with an alpha-blocker and low-dose anticholinergic combination therapy may be a reasonable and effective therapeutic option as an initial therapy.39 The relative benefit of anticholinergics compared to alpha-blocker only in terms of emptying efficiency and storage symptoms was retrospectively studied. One hundred and sixty-eight male LUTS patients with more than 8 IPSS score and more than 2 urgency score were enrolled.

furfur (76.56%), followed by M. sympodialis (12.50%) and M. japonica (9.38%). The most frequently isolated species in healthy individuals were M. furfur (61.67%), followed by M. sympodialis (25.00%), M. japonica

(6.67%), M. globosa (3.33%), and M. obtusa (3.33%). Overall, our study revealed that while M. furfur is the predominant Malassezia species in Chinese SD patients, there is no significant difference in the distribution of Malassezia species between Chinese SD patients and healthy individuals. “
“Critically ill patients admitted to intensive care units (ICU) are highly susceptible to healthcare-associated infections Histone Methyltransferase inhibitor caused by fungi. A prospective sequential survey of invasive fungal infections was conducted from May 2006 to April 2008 in 38 ICUs of 27 Italian hospitals. A total of 384 fungal infections (318 invasive Candida infections, three cryptococcosis and 63 mould infections) were notified. The median rate of candidaemia was 10.08 per 1000 admissions. In 15% of cases, the infection was already present at the time of admission to ICU. Seventy-seven percent of Candida infections were diagnosed in surgical patients. Candida albicans was isolated in 60% of cases, Candida glabrata and Candida parapsilosis in 13%, each. Candida glabrata had the

highest crude mortality rate (60%). Aspergillus infection was diagnosed in 32 medical and 25 surgical patients. The median rate was 6.31 per 1000 admissions. Corticosteroid treatment was the major host factor. Aspergillosis was demonstrated to be more severe than click here candidiasis as the crude mortality rate was significantly higher (63% vs. 46%), given an equal index of severity, Simplified Acute Physiology Score (SAPS-II). The present large nationwide

survey points out the considerable morbidity those and mortality of invasive fungal infections in surgical as well as medical patients in ICU. “
“Candida dubliniensis is a recently described yeast that causes infections in mucosal surfaces as well as sterile body sites. Candida dubliniensis develops resistance to fluconazole (FLC) more rapidly than the closely related species C. albicans. The killing activity of amphotericin B (AMB), 5-fluorocytosine (5FC), FLC, voriconazole (VRC) and posaconazole (POS) was determined against six C. dubliniensis clinical isolates, identified using molecular biological methods and C. dubliniensis CD36 reference strain. Minimum inhibitory concentrations (MICs) were determined using the Clinical and Laboratory Standards Institute standard procedure. Time-kill assays were performed using RPMI-1640 as test media over a 48-h period. AMB proved to be fungicidal at ≥0.5 μg ml−1 against all clinical isolates after 48 h. 5FC was only fungicidal at 32–64× MIC (4–8 μg ml−1) against all C. dubliniensis isolates. FLC, VRC and POS were fungistatic; decrease in colony number was observed only at the highest concentrations tested (8, 4 and 4 μg ml−1, respectively).

presents in healthy subjects, and Malassezia (5%) — which represents a twofold increase over healthy samples. In addition to the basiomycete fungi of the genus Cryptococcus, healthy scalps PARP inhibitor were dominated by Acremonium spp. and Didymella bryoniae (over 95% of the Ascomycota) [106]. An exemplary recent publication [79] has added further fundamental understanding of the role of skin microbiota in activating and educating

host immunity, shedding new light on the interplay between the immune system and microbiota. The authors studied patients with hyper IgE syndrome, a primary immunodeficiency resulting from STAT3 deficiency, and compared the bacterial and fungal skin microbiota at four clinically relevant sites PS-341 order representing the major skin microenvironments (the nares, retroauricular crease, antecubital fossa, and volar forearm) [79]. The patients displayed increased ecological permissiveness, characterized by altered microbial population

structures including colonization with bacterial microbial species not observed in healthy individuals, such as Clostridium species and Serratia marcescens [79]. An elevated fungal diversity and increased representation of opportunistic fungi (Candida and Aspergillus) were observed in hyper IgE syndrome patients, concomitant with a decrease in the relative abundance of the common skin fungus Malassezia [79]. These changes supported the hypothesis of increased skin permissiveness

Ribonucleotide reductase to microbial transit, suggesting that skin may serve as a reservoir for the recurrent fungal infections observed in these patients [79]. The differences in the cutaneous microbiota between healthy individuals and primary immunodeficiency patients probably correlate with their immunological status. Defects in STAT3 signaling impair defensin expression and the generation and recruitment of neutrophils [107], in part due to defects in Th17-cell differentiation. These findings further suggest that altered immune responses in disease modify not only the bacterial microbiota niche but also the fungal skin/mucosal communities, which may contribute to the increased fungal infections observed clinically in this patient population. The skin microbiota investigation provides an important step toward understanding the interactions between pathogenic and commensal fungal and bacterial communities, and how these interactions can result in beneficial or detrimental (i.e., disease) outcomes. Species often considered “normal” colonizers of the skin, such as Malassezia, can become causal agents of skin diseases. These preliminary results indicate the difficulty of defining a “normal” microbiota and consequently, meaningfully linking the mycobiota with clinical status would require a significant increase in the number of samples analyzed. The oral microbiota is a critical component of health and disease.

J.L.). The authors declare no financial or commercial conflict of interest. “
“There is a wealth of immunologic studies that have been carried out in experimental and human schistosomiasis that can be classified into three main areas: immunopathogenesis, resistance to reinfection and diagnostics. It is clear that the bulk of, if

not all, morbidity due to human schistosomiasis results from immune-response-based inflammation against eggs lodged in the body, either as regulated chronic inflammation or high throughput screening resulting in fibrotic lesions. However, the exact nature of these responses, the antigens to which they are mounted and the mechanisms of the critical regulatory responses are still being sorted out. It is also becoming

apparent that protective immunity against schistosomula as they develop into adult worms develops slowly and is hastened by the dying of adult worms, either naturally or when they are killed by praziquantel. However, as with anti-egg responses, the responsible immune mechanisms and inducing antigens are not clearly established, nor are any potential regulatory responses known. Finally, a wide variety of immune markers, both cellular and humoral, can be used to demonstrate exposure to schistosomes, and immunologic measurement of schistosome antigens can be used to detect, and thus diagnose, active infections. All three areas contribute to the public health response to human schistosome infections. Y-27632 clinical trial “
“Succinatimonas  hippei  is a new bacterial species isolated from human feces. Here we report that the growth of S. hippei YIT 12066T depends on CO2 or bicarbonate and the headspace gas produced by microbiota. Genetic defect for carbonic anhydrase in this bacterium suggested a reason for the syntrophic property of CO2 dependency and may suggest an adaptation to its habitat. The use of

culture-independent molecular methods to analyze gastrointestinal (GI) microbiota Aspartate has allowed more complete and accurate assessment of biodiversity in this ecosystem (1,2). Molecular methods using small subunit ribosomal RNA (SSU rRNA)-based technologies are considered useful for finding potential links between microbes and disease status. However, the results obtained with such approaches should not be considered to be suggestive of anything beyond microbial diversity, as potential functions of microbes cannot always be extracted from SSU rRNA data. To better understand the physiological characteristics and functions of the majority of human GI microbiota, we have been performing several intensive cultivation trials aimed at isolating so-called ‘unculturable’ or ‘as-yet-uncultured’ bacteria from the human GI tract (3–12). To date, we have isolated 17 new species of strictly anaerobic bacteria, including four new genera and two new families.