Short-term dietary adjustment with a hydrolyzed casein-based diet postpones diabetes development in the diabetes-prone BB rat

From earlier studies it appears that weaning associated changes in the animal's physiology and that of the pancreas in particular, render diabetes-prone Bio-Breeding (DP-BB) rats susceptible to the induction and development of insulin-dependent diabetes mellitus (IDDM). In this study we tested whether a short-term dietary adjustment at weaning would influence the development of diabetes later in life. For this purpose a diet in which the protein source was replaced with hydrolyzed casein (HC) was given to the rats from weaning to 60 days of age and from weaning to 130 days of age. The control group received the cereal-based standard diet throughout the experiment. The short-term dietary adjustment resulted in a significant delay of diabetes development. The rats fed the HC diet from weaning to 130 days of age showed a lower incidence of diabetes at 130 days of age. No differences were seen in the histological insulitis scores between the rats of the different treatment groups. Interestingly, when testing (mucosal) immune functions of short-term HC-fed rats, their mesenteric lymph node cells (MLNC) showed increased interferon-gamma (IFN-gamma) and reduced interleukin-10 (IL-10) production after in vitro stimulation. These results demonstrate that short-term dietary adjustments at a young age can influence the course of diabetes later in life. The shift in cytokine profile of MLNC of the HC-fed rats suggests that mechanisms involved can be at the level of both the (mucosal) immune system and the beta cell.     

Endoscopic Management of Infected Necrotizing Pancreatitis: an Evidence-Based Approach

Purpose of Review

Endoscopic management of infected necrotizing pancreatitis has evolved rapidly over the past years and there have been interesting innovations in this field. This review provides an update on the most recently published literature regarding endoscopic management of infected necrotizing pancreatitis.

Recent Findings

A recent randomized trial demonstrated no difference in mortality and major morbidity between endoscopic and surgical step-up treatment of infected necrotizing pancreatitis. However, endoscopic therapy resulted in shorter hospital stay and less pancreatic fistulas. Various innovations have been investigated with the aim to further optimize endoscopic therapy, in particular lumen-apposing metal stents. While major stent-related complications were also reported, findings from recent studies indicated that their use was associated with higher resolution rates of walled-off necrosis compared to double-pigtail stents. Other innovations, such as the multiple gateway technique and dual-modality mode, can be considered for treatment of particular cases. Furthermore, research suggests that irrigation of walled-off necrosis can be performed by using a nasocystic tube and discontinuation of proton-pump inhibitors may be considered.

Summary

Endoscopic treatment should be the preferred treatment modality in patients with infected necrotizing pancreatitis who are eligible for endoscopic drainage. Although data suggests that lumen-apposing metal stents are superior to double-pigtail stents, prospective multicenter studies focusing on safety as well as long-term follow-up are first needed.

     

Peptidoglycan-binding protein TsaP functions in surface assembly of type IV pili

Type IV pili (T4P) are ubiquitous and versatile bacterial cell surface structures involved in adhesion to host cells, biofilm formation, motility, and DNA uptake. In Gram-negative bacteria, T4P pass the outer membrane (OM) through the large, oligomeric, ring-shaped secretin complex. In the β-proteobacterium Neisseria gonorrhoeae, the native PilQ secretin ring embedded in OM sheets is surrounded by an additional peripheral structure, consisting of a peripheral ring and seven extending spikes. To unravel proteins important for formation of this additional structure, we identified proteins that are present with PilQ in the OM. One such protein, which we name T4P secretin-associated protein (TsaP), was identified as a phylogenetically widely conserved component of the secretin complex that co-occurs with genes for T4P in Gram-negative bacteria. TsaP contains an N-terminal carbohydrate-binding lysin motif (LysM) domain and a C-terminal domain of unknown function. In N. gonorrhoeae, lack of TsaP results in the formation of membrane protrusions containing multiple T4P, concomitant with reduced formation of surface-exposed T4P. Lack of TsaP did not affect the oligomeric state of PilQ, but resulted in loss of the peripheral structure around the PilQ secretin. TsaP binds peptidoglycan and associates strongly with the OM in a PilQ-dependent manner. In the δ-proteobacterium Myxococcus xanthus, TsaP is also important for surface assembly of T4P, and it accumulates and localizes in a PilQ-dependent manner to the cell poles. Our results show that TsaP is a novel protein associated with T4P function and suggest that TsaP functions to anchor the secretin complex to the peptidoglycan.Type IV pili systems (T4PSs) are involved in the assembly of long, thin fibers, which are found on the surfaces of many bacteria and archaea (1). Type IV pili (T4P) function in host cell adhesion, twitching motility, virulence, DNA uptake, and biofilm formation and are evolutionary related to type II secretion systems (T2SSs), bacterial transformation systems, and the archaellum (2–4). T4PSs can be divided into T4aPSs and T4bPSs that are distinguished based on pilin size and assembly systems (5, 6). T4aPSs form the most abundant class, and the T4P formed by these systems can undergo cycles of extension, adhesion, and retraction, which is a feature that distinguishes them from the other bacterial surface structures (7, 8). T4aP retract at rates up to 1 μm/s and can generate forces up to 150 pN (9, 10). Generally, T4bPSs are not associated with retraction. Here, we focus on T4aPSs and refer to these as T4PSs unless specifically indicated. T4PSs have been studied extensively in many bacteria but are especially well characterized in Neisseria and Pseudomonas spp. and in Myxococcus xanthus. Different nomenclature is used for different T4PSs (Table S1). Here, the Neisseria gonorrhoeae nomenclature is used.T4P are composed of major (e.g., PilE) and minor (in N. gonorrhoeae; e.g., PilV, PilX, ComP) pilins that are synthesized as preproteins with a type III signal peptide. After cleavage of the signal peptide by the prepilin peptidase PilD (11, 12), the T4P are assembled by a multiprotein complex (13). In Gram-negative bacteria, the proteins of T4PSs can be divided into three subcomplexes: the inner membrane (IM) motor complex, the alignment complex, and the outer membrane (OM) pore complex (6). The IM motor complex drives both the assembly and the retraction of T4P. Pilin subunits are extruded from the IM by the platform protein PilG (14) and the hexameric ATPase PilF (15). Disassembly of T4P with retraction occurs when PilF is replaced by the hexameric ATPase PilT (7, 16). PilU, a PilT paralog, is involved in retraction to a lesser extent (17). The alignment complex consisting of PilM, PilN, PilO, and PilP is proposed to connect the IM motor complex and the OM pore complex, and it is also thought to be involved in the stability and/or gating of the OM complex (18–20). In the OM, PilQ forms a homooligomeric ring that serves as a conduit for T4P (21–23).PilQ is a member of the secretin protein family. Proteins belonging to this family are present in many Gram-negative bacteria and are components of T4PSs, T2SSs, type III secretion systems (T3SSs), and extrusion systems of filamentous phages (24). Secretins are multidomain proteins with a signal sequence and a conserved C-terminal OM-spanning domain. Most secretins contain multiple copies of an N-terminal α/β domain (the N domains). PilQ proteins are integral OM proteins and form large gated channels. Oligomeric secretin complexes with different symmetries have been identified. Structural characterization by EM of purified PilQ from Neisseria meningitidis showed a dodecameric structure with a chamber sealed at both ends (25, 26), whereas the T2SS secretins PulD (27) and GspD (28) of the Klebsiella oxytoca pullanase and Vibrio cholerae toxin secretion systems, respectively, showed dodecameric structures with a chamber open at the periplasmic side and closed at the OM side. The structure of the InvG secretin complex of the T3SS of the Salmonella typhimurium needle complex showed 15-fold symmetry and is open at both ends (29), and the phage pIV secretin showed 14-fold symmetry (30). The structure of the C-terminal OM-spanning domain involved in multimer formation is currently not known. Crystal structures of the periplasmic N domains of GspD of the T2SS of enterotoxigenic Escherichia coli (31), of EscC of the T3SS of S. typhimurium (32), and of N. meningitidis PilQ (25) showed that these domains consist of α-helices packed against three-stranded β-sheets. Secretins of T4P systems also contain B domains, which are not present in other secretins and are located N-terminal to the N domains. The structure of the B2 domain of N. meningitidis PilQ consists of several β-strands (25). Remarkably, when the sequence conservation of the B2 domain was mapped to the structure of the B2 domain of N. meningitidis PilQ, a highly conserved patch was identified that was proposed to form the binding site for a currently unidentified T4PS protein (25).Secretins interact with several other proteins. Pilotin proteins are small lipoproteins that interact with the extreme C terminus of secretins and are responsible for OM targeting and oligomerization of secretins (33–38). Secretins of T4PSs also interact with the alignment complex. For N. meningitidis, Pseudomonas aeruginosa, and M. xanthus PilQ, a direct interaction was demonstrated between the respective PilPs and the N0 domains of the PilQs (25, 39, 40). Recently, ExeA of the T2SS of Aeromonas hydrophila (41) and FimV of the T4PS of P. aeruginosa (42) were also implicated in secretin assembly. They contain, respectively, PF01471 and LysM peptidoglycan (PG)-binding domains that might attach them to the PG. However, neither of these two proteins is ubiquitously conserved in bacteria assembling T4P.We have previously shown that the PilQ secretin of N. gonorrhoeae embedded in OM sheets is surrounded by a peripheral structure, which is formed by an additional peripheral ring as well as spikes (43). The proteins that make up these structures are not known. Here, we identify a widely conserved protein, which we name T4P secretin-associated protein (TsaP), that is important for the formation of the peripheral structure. Phylogenomic analysis of 450 genomes of Proteobacteria showed that the presence of the tsaP gene is strongly linked to the presence of genes for T4aPSs. We characterize the TsaP protein and demonstrate the importance of TsaP for T4aP assembly in the two phylogenetically widely separated model organisms N. gonorrhoeae and M. xanthus.     

Measurement properties of handheld dynamometry for assessment of shoulder muscle strength: A systematic review

Like any assessment tool, handheld dynamometry (HHD) must be valid and reliable in order to be meaningful in clinical practice and research. To summarize the evidence of measurement properties of HHD for the assessment of shoulder muscle strength. Cochrane Central Register of Controlled Trials (CENTRAL), PubMed, EMBASE, and PEDro were searched up to February 2020. Inclusion criteria were studies (a) evaluating HHD used on the glenohumeral joint, (b) evaluating measurement properties, and (c) included individuals ≥ 18 years old with or without shoulder symptoms. Exclusion criteria were studies (a) including patients with neurologic, neuromuscular, systemic diseases, or critical illness or bed-side patients and (b) that did not report the results separately for each movement. In total, 28 studies with 963 participants were included. The reliability results showed that 98% of the intraclass correlation coefficient (ICC) values were ≥0.70. The measurement error showed that the minimal detectable change in percent varied from 0% to 51.0%. The quality of evidence was high or moderate for the majority of movements and type of reliability examined. Based on the evidence of low or very low quality of evidence, the convergent validity and discriminative validity of HHD were either sufficient, indeterminate, or insufficient. The reliability of HHD was overall sufficient, and HHD can be used to distinguish between individuals on the group level. The measurement error was not sufficient, and evaluation of treatment effect on the individual level should be interpreted with caution.     

Allorecognition of HLA-DP by CD4+ T cells is affected by polymorphism in its alpha chain

Alloreactivity to HLA-DP molecules, class II heterodimers of an oligomorphic alpha and a polymorphic beta chain, is increasingly being studied due to its relevance in clinical transplantation. We hypothesized that not only polymorphisms in the peptide binding groove encoded by exon 2 of HLA-DPB1, but also in other regions of the molecule and the alpha chain, could play a role in CD4+ T cell allorecognition. To test this possibility, we comparatively investigated CD4+ T cell allorecognition, measured by upregulation of the activation marker CD137, against HLA-DPB1*13:01, *05:01, *03:01, *17:01 or their allele counter parts DPB1*107:01, *135:01, *104:01, *131:01, with identical exon 2 sequences but polymorphism in exons 1, 3 or 4, in the context of different HLA-DPA1 (DPA1) polymorphisms (DPA1*01:03 and *02:01). No significant differences in CD4+ T cell allorecognition levels could be demonstrated for any of the beyond exon 2 DPB1 variants studied. Interestingly, however, the mean fold change in CD4+ CD137+ cells was significantly higher when the target shared at least one DPA1 allele with the allogeneic stimulator, compared to a distinct DPA1 background (1.65 vs 0.23, P < 0.005). Structural homology modeling suggested specific amino acid residues in the alpha chain, in particular position 31, to impact CD4+ T cell allorecognition of HLA-DP. Our data argue against a significant role of beyond exon 2 DPB1 polymorphisms for T cell alloreactivity, but show relevance of DPA1 polymorphism in this mechanism. These new findings impact HLA matching strategies in unrelated stem cell transplantation.     

Monitoring CD8a+ T Cell Responses to Radiotherapy and CTLA-4 Blockade Using [64Cu]NOTA-CD8a PET Imaging

Molecular Imaging and Biology - Current response assessment systems for cancer patients receiving immunotherapy are limited. This is due to the associated inflammatory response that may confound...     

Fourier analysis of video-keratographic data. A tool for separation of spherical, regular astigmatic and irregular astigmatic corneal power components

Topographic analysis of the cornea and computation of changes in corneal topography is difficult because of the lack of a standardized concept for analysing topographic information. We have used Fourier series harmonic analysis to decompose the often very complex information given in topographic images (TMS-1 video-keratograph). The polar data values arising from each mire were separated into spherical (average power), regular astigmatic (2′ harmonic) and non-regular astigmatic components. The non-regular astigmatic components were further divided into a decentration component (1′ harmonic) and a higher order irregular refractive component (root mean square (r.m.s.) of the residual between original data and the sum of spherical, 1′ and 2′ order components). Algorithms for utilizing the Fourier analysis technique on subjective refractive and on keratometric changes in regular astigmatism were also derived. Averages of the separate refractive components were calculated for groups of normal (right and left eye of 25 patients), keratoconic (13 eyes), and transplanted eyes (20 eyes), and average topographic maps were reconstructed and imaged. Changes in the separate topographic refractive components were compared with subjective refractive changes in eyes that had undergone combined myopic-astigmatic excimer laser ablation (8 eyes). Fourier series analysis was found superior to conventional keratometry in predicting spherical and regular astigmatic changes in subjective refraction. Fourier series analysis seems to be a powerful tool to decompose the information of complex topographic corneal images into rational optical components.     

VerstÄrkung der radiotherapeutischen wirkung auf HeLa-Zellen durch gemcitabine

BACKGROUND: Gemcitabine (2'.2'-difluorodeoxycytidine; dFdC) is a new nucleoside analog with promising activity in different solid tumors in vivo and in vitro. As published up to now, combined with irradiation dFdC demonstrates a radiosensitizing effect on pancreas and colon carcinoma cell lines. We investigated the influence of dFdC on the radiosensitization of human squamous carcinoma cells of the cervix (HeLa-cells, ATCC CCL-2). MATERIAL AND METHODS: Under standardized conditions monolayer cultures of HeLa-cells were incubated in medium with dFdC for different times (4 to 24 hours) and exposed to different concentrations (0.003, 0.01 and 0.03 mumol/l). Irradiation (2 to 6 Gy, electron beam) followed immediately or 12 hours after dFdC-exposure. Cell survival was determined by colony forming assay. Using the linear-quadratic model cell survival curves were fit after correction for drug-induced cytotoxicity and the mean inactivation dose (MID) was calculated. Radiation enhancement was defined as the ratio MIDRT(= Control)/MIDRT + dFdC > 1. RESULTS: Exposed to gemcitabine for 4 and 8 hours and followed by immediate irradiation the radiation enhancement ratio (Table 1) is 1.07 to 1.14 and 1.04 to 1.22, respectively, if dFdC concentration is > or = 0.01 to 0.03 mumol/l. Further increase of the irradiation effect is demonstrated in cells exposed to > or = 0.003 to 0.03 mumol/l dFdC for 16 and 24 hours (radiation enhancement ratio 1.08 to 2.0 and 1.08 to 2.48, respectively) (Figure 3). If irradiation is applied 12 hours after 24-hour-exposure (0.01 and 0.03 mumol/l) the enhancement ratio was 1.18 and 1.7, respectively (Figure 4). CONCLUSIONS: In cell cultures the assays combining irradiation with dFdC demonstrate that dFdC is a potent radiation sensitizer of HeLa-cells. The effect of irradiation on cells pre-treated with non- and hardly cytotoxic concentrations of dFdC is increased in dependence of dose and time of exposure.