Impact of dietary deviation on disease progression and gut microbiome composition in lupus-prone SNF1 mice.

Environmental factors, including microbes and diet, play a key role in initiating autoimmunity in genetically predisposed individuals. However, the influence of gut microflora in the initiation and progression of systemic lupus erythematosus (SLE) is not well understood. In this study, we have examined the impact of drinking water pH on immune response, disease incidence and gut microbiome in a spontaneous mouse model of SLE. Our results show that (SWR × NZB) F1 (SNF1 ) mice that were given acidic pH water (AW) developed nephritis at a slower pace compared to those on neutral pH water (NW). Immunological analyses revealed that the NW-recipient mice carry relatively higher levels of circulating autoantibodies against nuclear antigen (nAg) as well as plasma cells. Importantly, 16S rRNA gene-targeted sequencing revealed that the composition of gut microbiome is significantly different between NW and AW groups of mice. In addition, analysis of cytokine and transcription factor expression revealed that immune response in the gut mucosa of NW recipient mice is dominated by T helper type 17 (Th17) and Th9-associated factors. Segmented filamentous bacteria (SFB) promote a Th17 response and autoimmunity in mouse models of arthritis and multiple sclerosis. Interestingly, however, not only was SFB colonization unaffected by the pH of drinking water, but also SFB failed to cause a profound increase in Th17 response and had no significant effect on lupus incidence. Overall, these observations show that simple dietary deviations such as the pH of drinking water can influence lupus incidence and affect the composition of gut microbiome.

Gender bias in lupus: does immune response initiated in the gut mucosa have a role?

The risk of developing systemic lupus erythematosus (SLE) is approximately nine times higher among women compared to men. However, very little is understood concerning the underlying mechanisms that contribute to this gender bias. Further, whether there is a link between immune response initiated in the gut mucosa, the progression of SLE and the associated gender bias has never been investigated. In this report, we show a potential link between the immune response of the gut mucosa and SLE and the gender bias of lupus for the first time, to our knowledge. Both plasma cell- and gut-imprinted- α4ß7 T cell frequencies were significantly higher in the spleen and gut mucosa of female (SWR × NZB)F1 (SNF1 ) mice compared to that of their male counterparts. Importantly, female SNF1 mice not only showed profoundly higher CD45(+) immune cell densities, but also carried large numbers of interleukin (IL)-17-, IL-22- and IL-9-producing cells in the lamina propria (LP) compared to their male counterparts. Intestinal mucosa of female SNF1 mice expressed higher levels of a large array of proinflammatory molecules, including type 1 interferons and Toll-like receptors 7 and 8 (TLR-7 and TLR-8), even before puberty. Our work, therefore, indicates that the gut immune system may play a role in the initiation and progression of disease in SLE and the associated gender bias.

Comparison of vestipitant with ondansetron for the treatment of breakthrough postoperative nausea and vomiting after failed prophylaxis with ondansetron.

BACKGROUND: Postoperative nausea and vomiting (PONV) is common; ondansetron is often used as prophylaxis or for breakthrough episodes. Vestipitant is a neurokinin 1 (NK-1) receptor antagonist that is effective for prophylaxis, but its efficacy for treating established PONV is unknown. This study was performed to evaluate the efficacy and safety of vestipitant, compared with ondansetron for the treatment of breakthrough PONV in patients who had already received prophylactic ondansetron before surgery. METHODS: A multicentre, randomized, single-blind (sponsor-open), parallel group study. Of 527 surgical patients, 130 (25%) had breakthrough PONV and were equally randomized to one of six i.v. doses of vestipitant (4-36 mg) or ondansetron 4 mg. The primary endpoint was the rate of patients exhibiting complete response, defined as no emesis and no further rescue medication from 10 min after infusion up to 24 h after surgery or hospital discharge. RESULTS: All doses of vestipitant were non-inferior to ondansetron in treating PONV after failed prophylaxis with ondansetron. However, vestipitant was superior to ondansetron in decreasing episodes of postoperative emesis and retching. The complete response rate analysis using Bayesian model averaging indicated that no vestipitant dose was superior to ondansetron. Nausea numerical rating scale scores and the times-to-PONV or discharge were similar between the vestipitant and ondansetron treatment groups. CONCLUSIONS: Although overall efficacy was non-inferior between vestipitant and ondansetron, the rate of emesis was lower with vestipitant. These data suggest that vestipitant may be a useful agent for the management of PONV, similar to other NK-1 antagonists. CLINICAL TRIAL REGISTRATION: NCT01507194.

Adrenal response to adrenocorticotropic hormone in dogs before and after surgical attenuation of a single congenital portosystemic shunt.

BACKGROUND: Dogs with single congenital portosystemic shunts (CPSS) often develop postoperative hypoglycemia and prolonged anesthetic recovery. These abnormalities could be attributable to inadequate adrenal response. However, adequacy of adrenal response after CPSS surgery is unexplored. HYPOTHESIS: Dogs with CPSS have inadequate postoperative adrenal response. ANIMALS: Eight nonoperated, 8 ovariohysterectomy (OHE), and 16 CPSS dogs. METHODS: Consecutive day ACTH stimulation tests were performed on nonoperated healthy dogs, healthy dogs before and after OHE, and CPSS dogs before and after surgery. Adequate response was defined as >50% or >30 ng/mL increase in cortisol after ACTH administration. Blood glucose (BG) was monitored before and after surgery. Prolonged anesthetic recovery and refractory hypoglycemia episodes were recorded. RESULTS: Results of consecutive day ACTH stimulation tests did not vary in normal dogs. Results of preoperative ACTH stimulation tests of CPSS and OHE dogs were not significantly different. Dogs with CPSS had higher postoperative baseline cortisol concentrations (median, 329 ng/mL) than OHE dogs (median, 153 ng/mL). Postoperative cortisol increase after ACTH in CPSS was < or =50% in 10/16 and < or =30 ng/mL in 6/16. After surgery, BG was < or =60 mg/dL in 7/16 CPSS dogs. Cortisol concentrations were not correlated with BG. Two CPSS dogs had refractory hypoglycemia and 4 had delayed recovery; all improved with dexamethasone administration (0.1-0.2 mg/kg/IV). CONCLUSIONS AND CLINICAL IMPORTANCE: Contrary to previous reports, baseline cortisol concentrations in CPSS and healthy dogs are similar. Many CPSS dogs have postoperative hypercortisolemia. Response to ACTH does not correlate with postoperative hypoglycemia or prolonged anesthetic recovery.

Side extraction duoPIGatron-type ion source.

We have designed and constructed a compact duoPIGatron-type ion source, for possible use in ion implanters, in which ions are extracted from a side aperture in contrast to conventional duoPIGatron sources with axial ion extraction. The size of the side extraction aperture is 1 x 40 mm(2). The ion source was developed to study physical and technological aspects relevant to an industrial ion source. The side extraction duoPIGatron has a stable arc, uniformly bright illumination, and dense plasma. The present work describes some operating parameters of the ion source using argon and BF(3). Total unanalyzed beam currents were 40 mA with Ar at an arc current of 7 A and 13 mA with BF(3) gas at an arc current of 9 A.

Bernas ion source discharge simulation.

As the technology and applications continue to grow up, the development of plasma and ion sources with clearly specified characteristic is required. Therefore comprehensive numerical studies at the project stage are the key point for ion implantation source manufacturing (especially for low energy implantation). Recently the most commonly encountered numerical approach is the Monte Carlo particle-in-cell (MCPIC) method also known as particle-in-cell method with Monte Carlo collisions. In ITEP the 2D3V numerical code PICSIS-2D realizing MCPIC method was developed in the framework of the joint research program. We present first results of the simulation for several materials interested in semiconductors. These results are compared with experimental data obtained at the ITEP ion source test bench.

Status of ITEP decaborane ion source program.

The joint research and development program is continued to develop steady-state ion source of decaborane beam for ion implantation industry. Both Freeman and Bernas ion sources for decaborane ion beam generation were investigated. Decaborane negative ion beam as well as positive ion beam were generated and delivered to the output of mass separator. Experimental results obtained in ITEP are presented.

Ion sources for energy extremes of ion implantation.

For the past four years a joint research and development effort designed to develop steady state, intense ion sources has been in progress with the ultimate goal to develop ion sources and techniques that meet the two energy extreme range needs of meV and hundreads of eV ion implanters. This endeavor has already resulted in record steady state output currents of high charge state of antimony and phosphorus ions: P(2+) [8.6 pmA (particle milliampere)], P(3+) (1.9 pmA), and P(4+) (0.12 pmA) and 16.2, 7.6, 3.3, and 2.2 pmA of Sb(3+)Sb(4+), Sb(5+), and Sb(6+) respectively. For low energy ion implantation, our efforts involve molecular ions and a novel plasmaless/gasless deceleration method. To date, 1 emA (electrical milliampere) of positive decaborane ions was extracted at 10 keV and smaller currents of negative decaborane ions were also extracted. Additionally, boron current fraction of over 70% was extracted from a Bernas-Calutron ion source, which represents a factor of 3.5 improvement over currently employed ion sources.

Determining incidence of extended spectrum beta-lactamase producing Enterobacteriaceae, vancomycin-resistant Enterococcus faecium and methicillin-resistant Staphylococcus aureus in 38 centres from 17 countries: the PEARLS study 2001-2002.

The PEARLS study prospectively monitored selected nosocomial pathogens from 38 centres in 13 European, three Middle Eastern countries and South Africa during 2001-2002. Extended spectrum beta-lactamase (ESBL) production rates among Escherichia coli, Klebsiella pneumoniae, and Enterobacter spp. were 5.4% (142/2609), 18.2% (401/2,206) and 8.8% (204/2,328), respectively, for all study sites. The overall ESBL production rate for the combined Enterobacteriaceae was 10.5% (747/7,143), highest in Egypt, 38.5%, and Greece, 27.4%, and lowest in The Netherlands, 2.0%, and Germany, 2.6%. IEF, PCR and DNA sequencing determined 10.7% false positives among Enterobacter spp. when using NCCLS guidelines to screen for ESBL production. The prevalence of nosocomial methicillin-resistant Staphylococcus aureus and vancomycin-resistant Enterococcus faecium was 32.4% (294/908) and 8.7% (83/949), respectively. PEARLS provides baseline data against which prospective changes in resistant determinants and outcomes can be measured in this ongoing study.

In vitro activity of gemifloxacin and contemporary oral antimicrobial agents against 27247 Gram-positive and Gram-negative aerobic isolates: a global surveillance study.

This study was a multi-centre, multi-country surveillance of 27247 Gram-positive and Gram-negative isolates collected from 131 study centres in 44 countries from 1997 to 2000. MICs of gemifloxacin were compared with penicillin, amoxicillin-clavulanic acid, cefuroxime, azithromycin, clarithromycin, trimethoprim-sulphamethoxazole, ciprofloxacin, grepafloxacin and levofloxacin by broth microdilution. Penicillin resistance in Streptococcus pneumoniae was extremely high in the Middle East (65.6%), Africa (64.0%) and Asia (60.4%) and lower in North America (40.3%), Europe (36.9%) and the South Pacific (31.8%). Macrolide resistance in S. pneumoniae was highest in Asia (51.7%) but varied widely between laboratories in Europe (26.0%), North America (21.6%), the Middle East (13.7%), the South Pacific (10.6%) and Africa (10.0%). All the study quinolones were highly active against penicillin-resistant and macrolide-resistant S. pneumoniae. Overall, gemifloxacin had the lowest MIC(90) at 0.06 mg/l with MICs 4-64-fold lower than ciprofloxacin, levofloxacin and grepafloxacin against S. pneumoniae. Gemifloxacin MICs were more potent than grepafloxacin > levoflaxacin > ciproflaxin against the Gram-positive aerobes and shared comparable Gram-negative activity with ciprofloxacin and levofloxacin.

Screening botanical extracts for quinoid metabolites.

Botanical dietary supplements represent a significant share of the growing market for alternative medicine in the USA, where current regulations do not require assessment of their safety. To help ensure the safety of such products, an in vitro assay using pulsed ultrafiltration and LC-MS-MS has been developed to screen botanical extracts for the formation of electrophilic and potentially toxic quinoid species upon bioactivation by hepatic cytochromes P450. Rat liver microsomes were trapped in a flow-through chamber by an ultrafiltration membrane, and samples containing botanical extracts, GSH and NADP(H), were flow-injected into the chamber. Botanical compounds that were metabolized to reactive intermediates formed stable GSH adducts mimicking a common in vivo detoxification pathway. If present in the ultrafiltrate, GSH conjugates were detected using LC-MS-MS with precursor ion scanning followed by additional characterization using product ion scanning and comparison to standard compounds. As expected, no GSH adducts of reactive metabolites were found in extracts of Trifolium pratense L. (red clover), which are under investigation as botanical dietary supplements for the management of menopause. However, extracts of Sassafras albidum (Nutt.) Nees (sassafras), Symphytum officinale L. (comfrey), and Rosmarinus officinalis L. (rosemary), all of which are known to contain compounds that are either carcinogenic or toxic to mammals, produced GSH adducts during this screening assay. Several compounds that formed GSH conjugates including novel metabolites of rosmarinic acid were identified using database searching and additional LC-MS-MS studies. This assay should be useful as a preliminary toxicity screen during the development of botanical dietary supplements. A positive test suggests that additional toxicological studies are warranted before human consumption of a botanical product.

The impact of P-glycoprotein efflux on enterocyte residence time and enterocyte-based metabolism of verapamil.

P-glycoprotein (P-gp) can limit the intestinal permeability of a number of compounds and may therefore influence their exposure to metabolizing enzymes within the enterocyte (e.g. cytochrome P450 3A, CYP 3A). In this study, the intestinal metabolic profile of verapamil, the influence of P-gp anti-transport on the cellular residence time of verapamil, and the impact of this change in residence time on the extent of enterocyte-based metabolism have been investigated in-vitro, utilizing segments of rat jejunum and side-by-side diffusion chambers. Verapamil exhibited concentration-dependent P-gp efflux and CYP 3A metabolism. The P-gp efflux of verapamil (1 microM) increased the cellular residence time across the intestinal membrane (approximately 3-fold) in the mucosal to serosal (m to s) direction relative to serosal to mucosal (s to m), yielding significantly greater metabolism (approximately 2-fold), presumably as a result of the prolonged exposure to CYP 3A. Intestinal metabolism of verapamil generated not only norverapamil, but resulted also in the formation of an N-dealkylated product (D-617). Norverapamil and D-617 accumulated significantly in mucosal chambers, relative to serosal chambers, over the time course of the experiment. Based on these in-vitro data, it was apparent that P-gp efflux prolonged the cellular residence time of verapamil (m to s) and therefore increased the extent of intestinal metabolism, and also played a role in metabolite secretion from within the enterocyte.

Combined c-fos and 14C-2-deoxyglucose method to differentiate site-specific excitation from disinhibition: analysis of maternal behavior in the rat.

On the basis of evidence that 14C-2-deoxyglucose (2-DG) autoradiography indicates activity at axonal terminals, whereas c-fos immunocytochemistry indicates activity of neuronal cell bodies, we combined these techniques in adjacent histological brain sections to assess excitatory and disinhibitory synaptic relations in selected sites in female rats in which maternal behavior was elicited by natural parturition, sensitization (7- to 10-day cohabitation with foster pups), or hysterectomy. All individuals in these three groups expressed maternal behavior immediately before 2-DG injection. Controls were non-maternal virgins. Parturient and Hysterectomized groups: elevation (compared with controls) in both 2-DG and c-fos activity in medial preoptic area (MPOA) indicated an increase in its input and output activity, i.e., an excitatory interaction; the MPOA was previously shown to be critical for maternal behavior. Sensitized group: a decrease in 2-DG activity of vomeronasal nuclei (bed nucleus of the accessory olfactory tract, BAOT, and medial amygdala, ME, replicating our previous study) and an elevation in c-fos activity, jointly indicate disinhibition of these nuclei, that were previously shown to modulate pup-chemostimulation-induced sensitization. All other sites showed evidence of excitatory input-output relationships (i.e., joint increase in both 2-DG and c-fos activity), e.g., bed nucleus of the stria terminalis (BNST), lateral habenula (LHAB), central gray (CG), thalamus (THAL), septum (SEPT), and ventral tegmental area (VTA). The present study demonstrates the feasibility of measuring 2-DG and c-fos activity jointly in adjacent sections of the same brain, thereby providing evidence to distinguish between localized excitation and disinhibition.

Antinociceptive action of vaginocervical stimulation in rat spinal cord: 2-DG analysis.

Using [14C] 2-deoxyglucose (2-DG) autoradiography with computerized densitometric analysis, unilateral foot pinch was found to significantly increase the relative optical density in laminae I and II of the ipsilateral, compared to the contralateral, spinal cord at lumbar 5 (L5). However, during vaginocervical mechanostimulation applied concurrently with the unilateral foot pinch, no comparable difference was observed. No response to foot pinch was observed in other laminae of the spinal cord at L5, and no effects comparable to the above were observed at L3. These findings indicate that vaginocervical mechanostimulation suppresses neural responses to noxious foot pinch stimulation selectively at the laminae I and II level of the spinal cord at L5, but not at L3.

Brain 2-deoxyglucose levels related to maternal behavior-inducing stimuli in the rat.

Levels of [14C]2-deoxyglucose (2-DG), measured autoradiographically, in the medial preoptic area (MPOA), were higher during natural parturition with concurrent maternal behavior than in non-pregnant non-maternal controls, whereas levels in the vomeronasal system were lower in virgin rats made maternal by cohabitation with young than in control and parturient rats. Previous studies have shown that lesions of MPOA disrupt maternal behavior, whereas lesions of vomeronasal structures stimulate it, and that an increase in 2-DG levels is indicative of an increase in firing activity in neuron terminals. Consequently, the present findings suggest that maternal behavior can be induced by: (a) an increase in parturition-generated sensory stimulatory input to the MPOA in response to mechanostimulation of the birth canal, and (b) a separate chemosensory vomeronasal pathway whose activity is reduced cohabitation with young, thereby disinhibiting maternal behavior.

The natural history of the anterior cruciate ligament-deficient knee. A review.

The ACL-deficient knee has been a management dilemma for many years and, to this day, no refutable plan exists for treatment of this injury. No true prospective study has been performed that evaluates all types of individuals at a variety of activity levels, and, in this day of apparently reliable methods of reconstructing the ACL, it is doubtful that one will occur. The ACL injury is no longer a mystery to the general public; it has received extensive publicity because of injuries of professional athletes and the successful reconstruction in many of these athletes. This article has not completely cleared up the issue of the future of an ACL-deficient knee. It has provided, however, convincing evidence that an active individual with a nonfunctional ACL is susceptible to meniscus injury (R. Barrack, J. Bruckner, J. Kneisl, et al, personal communication, 1990). There is also the risk of more tears occurring with time. Bray and Dandy found in their follow-up of patients with ACL repairs that, if the pivot shift returned, these patients had a much higher incidence of meniscus tears. Many of these studies indicate that, if the meniscus cannot be repaired and requires partial meniscectomy or worse, the articular surface will deteriorate (R. Barrack, J. Bruckner, J. Kneisl, et al, personal communication, 1990). Satku et al showed only 11% incidence of radiographic changes in patients with ACL-deficient knees with no evidence of meniscus tears compared with 100% in those having meniscectomy more than 5 years previously. Activity levels in general also change following this injury. This is probably the most difficult area to assess. Even though a substantial number of persons returned to their preinjury level of activity, it is not always possible to determine if they are playing with the same behavior and attitude. In other words, athletes who are involved in sports with cutting and jumping may modify the need for these activities and yet remain relatively competitive depending on their previous level of skill and the position they play. It has also been shown that many athletes return to their preinjury level initially but with time have significant increase in their symptoms and must modify their level of participation. More individuals limited their activities from the beginning than returned to their preinjury level (R. Barrack, J. Bruckner, J. Kneisl, et al, personal communication, 1990). Instability varies in these individuals and, as in Chick and Jackson's patients, those with mild instability (no rotatory instability) may do reasonably well.(ABSTRACT TRUNCATED AT 400 WORDS)

Ultrastructural and paraphenylene studies of degeneration in the primate visual system: degenerative remnants persist for much longer than expected.

It is a widely held belief that the products of axonal degeneration in the CNS are transitory and are caused by metabolic and phagocytic processes. However, recent light microscopic examinations of human and primate brains using the paraphenylene diamine staining method (PPD), which stains degenerating axons, have confirmed that the products of degeneration persist for years in visual pathways. The routine utilization of the PPD method for delineating human visual pathways requires further confirmation of axonal degeneration. Optic nerves, optic tracts, and lateral geniculate nuclei were collected from human brains that had clinical documentation of optic nerve damage prior to death. Optic nerves, optic tracts, and lateral geniculate nuclei taken from the brains of cynomolgus monkeys that had undergone enucleation 3 months to 1 year prior to sacrifice were also examined. All tissue was processed for electron microscopy; ultrathin sections were cut for electron microscopy, and consecutive sections were cut for light microscopy. In all cases, the homology of the degenerated processes was confirmed between the light microscopic (PPD) and the electron microscopic sections. Such ultrastructural examination demonstrates that the products of axonal degeneration remain in the primate visual system longer than previously supposed.

Immune response to a thymus-dependent form of B512 dextran requires the presence of Lyb-5+ lymphocytes.

Studies of the ontogeny of the immune response to B512 dextran (Dex) show that antibody responses equal to those of adult mice are not attained until 12 wk of age. We have examined the anti-Dex response after immunization with a thymus-dependent antigen isomaltohexaosyl-keyhole limpet hemocyanin (IM6-KLH) and have shown that the development of the cross-reacting anti-Dex response parallels the development of Lyb-5+ B cells. Adult levels of anti-Dex antibody after immunization with IM6-KLH are achieved in mice between 3 and 12 wk of age, a time when Lyb-5+ cells have reached adult levels. Neonatal mice, immunized at 1 d or 1 wk after birth, failed to produce a significant amount of anti-Dex antibodies, although they did produce IM6-specific antibodies after immunization with IM6-KLH. Data, which support the conclusion from these experiments that Lyb-5+ cells are required for an anti-polysaccharide response even when the immunizing antigen is thymus-dependent, include the failure of IM6-KLH to stimulate a normal anti-Dex response in mice with the xid defect and the direct demonstration in normal adult mice that elimination of Lyb-5+ cells from spleens of mice primed with IM6-KLH abolishes the ability of these cells to transfer an anti-Dex response. The data imply that the expressed B cell repertoire in adult animals is skewed such that the vast majority of B cells capable of responding to polysaccharide determinants are in the Lyb-5+ subset.