Surgical Emergencies in Inflammatory Bowel Disease.

Inflammatory bowel disease (IBD) patients are at risk for undergoing emergency surgery for fulminant disease, toxic megacolon, bowel perforation, intestinal obstruction, or uncontrolled gastrointestinal hemorrhage. Unfortunately, medical advancements have failed to significantly decrease rates of emergency surgery for IBD. It is therefore important for all acute care and colorectal surgeons to understand the unique considerations owed to this often-challenging patient population.

Anal Cryptoglandular Suppuration: Evidence-Based Management.

Anal suppurative processes are commonly encountered in surgical practice. While the initial therapeutic intervention is philosophically straightforward (incision and drainage), drainage of the appropriate space and treatment of the subsequent fistula in ano require a thorough understanding of perianal anatomy and nuanced decision making. Balancing the risk of fecal incontinence with simple fistulotomy versus the higher risk of fistula recurrence with all sphincter-sparing fistula treatments can be a challenge for surgeons and patients alike.

Pathology caused by introduced Neoergasilus japonicus (Copepoda: Ergasilidae) to the skin of indigenous Tilapia sparrmanii in South Africa and scanning electron microscopy study of wound-inflicting structures.

Neoergasilus japonicus, a fish ectoparasite indigenous to eastern and southern Asia, has been introduced to various regions, including Europe and North and South America, where it spreads at an alarming rate. The parasite is not host-specific. It was also collected from the Vaal-Orange and Limpopo river systems in South Africa. This report describes the morphology of the wound-inflecting structures in N. japonicus and relates their morphology to the pathological effect on Tilapia sparrmanii. Host tissue with parasites attached was imbedded in resin for histological sectioning and stained with haematoxylin and eosin. Additional specimens were prepared for scanning electron microscopy. Adult female parasites attach by inserting the hooked tips of the antennae into the epidermis. The maxillulae are minute but have sharp tips. The maxillae have sharp tips on the endopod and brush-like setae on the exopod. The mandibles are armed with sharp tips. These structures are used to remove the epidermis of the host and brush it into the buccal cavity. Host tissue in the intestine confirms that adult females consume host tissue leaving the dermis exposed to opportunistic diseases. The microscopic size of the parasite prevents casual observation, and its occurrence in Africa is, therefore, probably underreported.

Hierarchical design of pseudosymmetric protein nanoparticles.

Discrete protein assemblies ranging from hundreds of kilodaltons to hundreds of megadaltons in size are a ubiquitous feature of biological systems and perform highly specialized functions 1-3. Despite remarkable recent progress in accurately designing new self-assembling proteins, the size and complexity of these assemblies has been limited by a reliance on strict symmetry 4,5. Inspired by the pseudosymmetry observed in bacterial microcompartments and viral capsids, we developed a hierarchical computational method for designing large pseudosymmetric self-assembling protein nanomaterials. We computationally designed pseudosymmetric heterooligomeric components and used them to create discrete, cage-like protein assemblies with icosahedral symmetry containing 240, 540, and 960 subunits. At 49, 71, and 96 nm diameter, these nanoparticles are the largest bounded computationally designed protein assemblies generated to date. More broadly, by moving beyond strict symmetry, our work represents an important step towards the accurate design of arbitrary self-assembling nanoscale protein objects.

Hierarchical design of pseudosymmetric protein nanoparticles.

Discrete protein assemblies ranging from hundreds of kilodaltons to hundreds of megadaltons in size are a ubiquitous feature of biological systems and perform highly specialized functions1-3. Despite remarkable recent progress in accurately designing new self-assembling proteins, the size and complexity of these assemblies has been limited by a reliance on strict symmetry4,5. Inspired by the pseudosymmetry observed in bacterial microcompartments and viral capsids, we developed a hierarchical computational method for designing large pseudosymmetric self-assembling protein nanomaterials. We computationally designed pseudosymmetric heterooligomeric components and used them to create discrete, cage-like protein assemblies with icosahedral symmetry containing 240, 540, and 960 subunits. At 49, 71, and 96 nm diameter, these nanoparticles are the largest bounded computationally designed protein assemblies generated to date. More broadly, by moving beyond strict symmetry, our work represents an important step towards the accurate design of arbitrary self-assembling nanoscale protein objects.

Genetic characterisation of four Lamproglena spp. (Copepoda, Lernaeidae) from Africa and the first mitochondrial data.

Females of species of Lamproglena von Nordmann, 1832 are parasitic on the gills of teleost fishes and the 38 nominal species are based on mainly morphological data. Only four of these species have been genetically characterised and no mitochondrial data are available for the genus. The present study aimed to provide representative ribosomal DNA (rDNA) data for two additional species of Lamproglena from Africa: Lamproglena clariae Fryer, 1956 and Lamproglena hoi Dippenaar, Luus-Powell et Roux, 2001, alongside mitochondrial DNA (mtDNA) for these and two other African species, Lamproglena hemprichii von Nordmann, 1832 and Lamproglena monodi Capart, 1944. The four species were collected from Clariidae, Cyprinidae, Alestidae and Cichlidae, respectively. Representative 18S rDNA and 28S rDNA data were obtained for L. clariae and L. hoi, while cox1 mtDNA was obtained for all four species. The respective haplotypes supported the distinctness of all species using all three gene regions investigated. Interestingly, species appeared to be grouped more by geographical origin than host family, with L. hoi more closely related to other African species than to Asian species also collected from cyprinid hosts. Even though the results presented here greatly add to the molecular data available for Lamproglena, there are still 32 (>80%) species for which no genetic data are available. The interpretation of the results presented here is thus preliminary and much more data are required before the phylogeny of this genus, and other members of the family, such as Lernaea Linnaeus, 1758, can be studied appropriately.

Computational design of constitutively active cGAS.

Cyclic GMP-AMP synthase (cGAS) is a pattern recognition receptor critical for the innate immune response to intracellular pathogens, DNA damage, tumorigenesis and senescence. Binding to double-stranded DNA (dsDNA) induces conformational changes in cGAS that activate the enzyme to produce 2'-3' cyclic GMP-AMP (cGAMP), a second messenger that initiates a potent interferon (IFN) response through its receptor, STING. Here, we combined two-state computational design with informatics-guided design to create constitutively active, dsDNA ligand-independent cGAS (CA-cGAS). We identified CA-cGAS mutants with IFN-stimulating activity approaching that of dsDNA-stimulated wild-type cGAS. DNA-independent adoption of the active conformation was directly confirmed by X-ray crystallography. In vivo expression of CA-cGAS in tumor cells resulted in STING-dependent tumor regression, demonstrating that the designed proteins have therapeutically relevant biological activity. Our work provides a general framework for stabilizing active conformations of enzymes and provides CA-cGAS variants that could be useful as genetically encoded adjuvants and tools for understanding inflammatory diseases.

Camallanid nematodes from Clarias gariepinus (Burchell, 1822) in the Crocodile River, Gauteng, South Africa: Exploring diversity and divergence in an acid-mine drainage impacted environment.

Clarias gariepinus collected from Lake Heritage, Crocodile River, were found to harbour camallanid nematodes. Previously, Boomker (1982) surveyed the Hartbeespoort Dam, downstream of the current study site, and identified a high prevalence of Procamallanus (Procamallanus) laeviconchus and Paracamallanus cyathopharynx. Since then, Procamallanus (Procamallanus) pseudolaeviconchus was described from C. gariepinus suggesting reconsideration of the identifications of Procamallanus species in historical studies from clariids. The aim of the current study was to definitively identify the nematodes collected from C. gariepinus in Lake Heritage, using morphological and molecular analyses. Morphological study consisted of light and scanning electron microscopy which confirmed the identity P. (P.) pseudolaeviconchus and P. cyathopharynx. This included descriptions of the detailed morphology of isolated buccal capsules for both species using soft tissue digestion, notably for the first time for P. (P.) pseudolaeviconchus. The morphology of isolated spiculae of both species was described for the first time using SEM. Molecular analyses included genetic characterisation of the small ribosomal subunit (18S) rDNA and cytochrome oxidase 1 (CO1) mtDNA. Genetic data supported the morphological identification of both species, however, divergence was detected in CO1 mtDNA data for P. cyathopharynx indicating two distinct lineages. Due to this variation, the morphometry of P. cyathopharynx specimens were revisited including statistical re-evaluation. No robust morphological traits were identified to support CO1 mtDNA lineages and all specimens were considered conspecific. In terms of camallanid biodiversity in the Crocodile River system, it is similar to that in Boomker (1982), despite the altered water quality from past acid mine pollution in the river.

The influence of signs of social class on compassionate responses to people in need.

A field experiment (N = 4,536) examined how signs of social class influence compassionate responses to those in need. Pedestrians in two major cities in the United States were exposed to a confederate wearing symbols of relatively high or low social class who was requesting money to help the homeless. Compassionate responding was assessed by measuring the donation amount of the pedestrians walking past the target. Pedestrians gave more than twice (2.55 times) as much money to the confederate wearing higher-class symbols than they did to the one wearing lower-class symbols. A follow-up study (N = 504) exposed participants to images of the target wearing the same higher- or lower-class symbols and examined the antecedents of compassionate responding. Consistent with theorizing, higher-class symbols elicited perceptions of elevated competence, trustworthiness, similarity to the self, and perceived humanity compared to lower-class symbols. These results indicate that visible signs of social class influence judgments of others' traits and attributes, as well as in decisions to respond compassionately to the needs of those who are suffering.

Acetylcholine-gated current translates wake neuronal firing rate information into a spike timing-based code in Non-REM sleep, stabilizing neural network dynamics during memory consolidation.

Sleep is critical for memory consolidation, although the exact mechanisms mediating this process are unknown. Combining reduced network models and analysis of in vivo recordings, we tested the hypothesis that neuromodulatory changes in acetylcholine (ACh) levels during non-rapid eye movement (NREM) sleep mediate stabilization of network-wide firing patterns, with temporal order of neurons' firing dependent on their mean firing rate during wake. In both reduced models and in vivo recordings from mouse hippocampus, we find that the relative order of firing among neurons during NREM sleep reflects their relative firing rates during prior wake. Our modeling results show that this remapping of wake-associated, firing frequency-based representations is based on NREM-associated changes in neuronal excitability mediated by ACh-gated potassium current. We also show that learning-dependent reordering of sequential firing during NREM sleep, together with spike timing-dependent plasticity (STDP), reconfigures neuronal firing rates across the network. This rescaling of firing rates has been reported in multiple brain circuits across periods of sleep. Our model and experimental data both suggest that this effect is amplified in neural circuits following learning. Together our data suggest that sleep may bias neural networks from firing rate-based towards phase-based information encoding to consolidate memories.

Morphological and molecular description of Allocreadium apokryfi sp. n. (Digenea: Allocreadiidae) from native Labeobarbus aeneus (Cyprinidae) in South Africa, including notes on its biology, evolutionary history and an updated key of African Allocreadium.

Adult trematodes of Allocreadium Looss, 1900 (Digenea) infect the intestine of mostly freshwater fishes in Asia, Europe, Africa and the Americas. During routine parasitological surveys in the Vaal River system, adult trematodes were collected from the intestine of smallmouth yellowfish, Labeobarbus aeneus (Burchell). The trematodes were confirmed to represent a member of Allocreadium and did not match any existing taxon. Therefore, they are described as a new species, Allocreadium apokryfi sp. n. The morphology of the new species most closely resembles that of Allocreadium aswanense El-Naffar, Saoud et Hassan, 1984, but it differs from it by having a bipartite internal seminal vesicle, wider eggs, a shorter intertesticular distance, an intestinal bifurcation at the ventral sucker level, a ventral sucker that is larger than the oral sucker, and a genital pore near the intestinal bifurcation or the ventral sucker. The surface topology of the new species is notably different from that of other allocreadiids. Papillae were observed in the ventral sucker and surrounding both ventral and oral suckers, but the number and arrangement of the latter were not consistent among specimens. The protruding cirrus of A. apokryfi sp. n. was described using SEM and is the first such observation for the genus. Genetic characterisation showed that the new species was clearly distinct from other Allocreadium spp. using both 18S (nucleotide difference 1.3-9.1%) and 28S (4.7-6.5%) rDNA, forming a well-supported clade in Allocreadium. The presence of A. apokryfi sp. n. in a well-studied river is unexpected, and considering the diet of its host and the scarcity of Allocreadium in Africa, the possible biology of this species is discussed herein.

Engineering a Plant-Derived Astaxanthin Synthetic Pathway Into Nicotiana benthamiana.

Carotenoids have been shown to be essential for human nutrition. Consumption of carotenoid-rich fruits and vegetables can reduce the risk of many diseases. The ketocarotenoid astaxanthin has become a commercially valuable compound due to its powerful antioxidant properties compared to other carotenoids. It is naturally produced in certain algae, bacteria, and the flowers of some species of the genus Adonis, although it is produced in such small quantities in these organisms that it is costly to extract. Chemical synthesis of this compound has also shown limited success with a high proportion of esterified forms of astaxanthin being produced, which decreases antioxidant properties by the conversion of hydroxyl groups to esters. Previously, transgenic astaxanthin-producing plants have been created using a ß-carotene ketolase enzyme of either bacterial or algal origin. However, a novel astaxanthin pathway exists in the flowering plants of the genus Adonis which has not been utilized in the same manner. The pathway involves two unique enzymes, ß-ring-4-dehydrogenase and 4-hydroxy-ß-ring-4-dehydrogenase, which add the necessary hydroxyl and ketone groups to the rings of ß-carotene. In the present study, Nicotiana benthamiana plants were transformed with chimeric constructs coding for these two enzymes. The regenerated, transgenic plants accumulate astaxanthin and their growth (height and weight) was unaffected, when compared to non-transformed N. benthamiana and to plants transformed with the bacterial ß-carotene ketolase. The accumulation of astaxanthin also improved seedling survivability under harsh UV light, mitigated reactive oxygen accumulation, and provided a phenotype (color) that allowed the efficient identification and recovery of transgenic plants with and without selection.

Early Resumption of ß Blockers Is Associated with Decreased Atrial Fibrillation after Noncardiothoracic and Nonvascular Surgery: A Cohort Analysis.

WHAT WE ALREADY KNOW ABOUT THIS TOPIC: WHAT THIS ARTICLE TELLS US THAT IS NEW: BACKGROUND:: Beta (ß) blockers reduce the risk of postoperative atrial fibrillation and should be restarted after surgery, but it remains unclear when best to resume ß blockers postoperatively. The authors thus evaluated the relationship between timing of resumption of ß blockers and atrial fibrillation in patients recovering from noncardiothoracic and nonvascular surgery. METHODS: The authors evaluated 8,201 adult ß-blocker users with no previous history of atrial fibrillation who stayed at least two nights after noncardiothoracic and nonvascular surgery as a retrospective observational cohort. After propensity score matching on baseline and intraoperative variables, 1,924 patients who did resume ß blockers by the end of postoperative day 1 were compared with 973 patients who had not resumed by that time on postoperative atrial fibrillation using logistic regression. A secondary matched analysis compared 3,198 patients who resumed ß blockers on the day of surgery with 3,198 who resumed thereafter. RESULTS: Of propensity score-matched patients who resumed ß blockers by end of postoperative day 1, 4.9% (94 of 1,924) developed atrial fibrillation, compared with 7.0% (68 of 973) of those who resumed thereafter (adjusted odds ratio, 0.69; 95% CI, 0.50-0.95; P = 0.026). Patients who resumed ß blockers on day of surgery had an atrial fibrillation incidence of 4.9% versus 5.8% for those who started thereafter (odds ratio, 0.84; 95% CI, 0.67-1.04; P = 0.104). CONCLUSIONS: Resuming ß blockers in chronic users by the end of the first postoperative day may be associated with lower odds of in-hospital atrial fibrillation. However, there seems to be little advantage to restarting on the day of surgery itself.

Appendiceal Neoplasms.

Appendiceal neoplasms are identified in 0.9 to 1.4% of appendiceal specimens, and the incidence is increasing. It has long been professed that neuroendocrine tumors (formerly carcinoids) are the most common neoplastic process of the appendix; recent data, however, has suggested a shift in epidemiology. Our intent is to distill the complex into an algorithm, and, in doing so, enable the surgeon to seamlessly maneuver through operative decisions, treatment strategies, and patient counseling. The algorithm for evaluation and treatment is complex, often starts from the nonspecific presenting complaint of appendicitis, and relies heavily on often subtle histopathologic differences.

Development of a thermostable nanoemulsion adjuvanted vaccine against tuberculosis using a design-of-experiments approach.

BACKGROUND: Adjuvants have the potential to increase the efficacy of protein-based vaccines but need to be maintained within specific temperature and storage conditions. Lyophilization can be used to increase the thermostability of protein pharmaceuticals; however, no marketed vaccine that contains an adjuvant is currently lyophilized, and lyophilization of oil-in-water nanoemulsion adjuvants presents a specific challenge. We have previously demonstrated the feasibility of lyophilizing a candidate adjuvanted protein vaccine against Mycobacterium tuberculosis (Mtb), ID93 + GLA-SE, and the subsequent improvement of thermostability; however, further development is required to prevent physicochemical changes and degradation of the TLR4 agonist glucopyranosyl lipid adjuvant formulated in an oil-in-water nanoemulsion (SE). MATERIALS AND METHODS: In this study, we took a systematic approach to the development of a thermostable product by first identifying compatible solution conditions and stabilizing excipients for both antigen and adjuvant. Next, we applied a design-of-experiments approach to identify stable lyophilized drug product formulations. RESULTS: We identified specific formulations that contain disaccharide or a combination of disaccharide and mannitol that can achieve substantially improved thermostability and maintain immunogenicity in a mouse model when tested in accelerated and real-time stability studies. CONCLUSION: These efforts will aid in the development of a platform formulation for use with other similar vaccines.

In vitro bioassessment of the immunomodulatory activity of Saccharomyces cerevisiae components using bovine macrophages and Mycobacterium avium ssp. paratuberculosis.

The yeast Saccharomyces cerevisiae and its components are used for the prevention and treatment of enteric disease in different species; therefore, they may also be useful for preventing Johne's disease, a chronic inflammatory bowel disease of ruminants caused by Mycobacterium avium ssp. paratuberculosis (MAP). The objective of this study was to identify potential immunomodulatory S. cerevisiae components using a bovine macrophage cell line (BOMAC). The BOMAC phagocytic activity, reactive oxygen species production, and immune-related gene (IL6, IL10, IL12p40, IL13, IL23), transforming growth factor ß, ARG1, CASP1, and inducible nitric oxide synthase expression were investigated when BOMAC were cocultured with cell wall components from 4 different strains (A, B, C, and D) and 2 forms of dead yeast from strain A. The BOMAC phagocytosis of mCherry-labeled MAP was concentration-dependently attenuated when BOMAC were cocultured with yeast components for 6 h. Each yeast derivative also induced a concentration-dependent increase in BOMAC reactive oxygen species production after a 6-h exposure. In addition, BOMAC mRNA expression of the immune-related genes was investigated after 6 and 24 h of exposure to yeast components. All yeast components were found to regulate the immunomodulatory genes of BOMAC; however, the response varied among components and over time. The in vitro bioassessment studies reported here suggest that dead yeast and its cell wall components may be useful for modulating macrophage function before or during MAP infection.

Functional network stability and average minimal distance - A framework to rapidly assess dynamics of functional network representations.

BACKGROUND: Recent advances in neurophysiological recording techniques have increased both the spatial and temporal resolution of data. New methodologies are required that can handle large data sets in an efficient manner as well as to make quantifiable, and realistic, predictions about the global modality of the brain from under-sampled recordings. NEW METHOD: To rectify both problems, we first propose an analytical modification to an existing functional connectivity algorithm, Average Minimal Distance (AMD), to rapidly capture functional network connectivity. We then complement this algorithm by introducing Functional Network Stability (FuNS), a metric that can be used to quickly assess the global network dynamic changes over time, without being constrained by the activities of a specific set of neurons. RESULTS: We systematically test the performance of AMD and FuNS (1) on artificial spiking data with different statistical characteristics, (2) from spiking data generated using a neural network model, and (3) using in vivo data recorded from mouse hippocampus during fear learning. Our results show that AMD and FuNS are able to monitor the change in network dynamics during memory consolidation. COMPARISON WITH OTHER METHODS: AMD outperforms traditional bootstrapping and cross-correlation (CC) methods in both significance and computation time. Simultaneously, FuNS provides a reliable way to establish a link between local structural network changes, global dynamics of network-wide representations activity, and behavior. CONCLUSIONS: The AMD-FuNS framework should be universally useful in linking long time-scale, global network dynamics and cognitive behavior.

Redescription of Cichlidogyrus philander (Monogenea, Ancyrocephalidae) using scanning electron microscopy (SEM) and molecular analysis.

The sclerotized structures of monogeneans have traditionally been studied by light microscopy and different staining techniques. Recently, enzymatic digestion followed by scanning electron microscopy (SEM) has enabled the examination of structural details not visible with light microscopy. In order to obtain better, and more accurate, morphological information on sclerotized structures not affected by mounting medium or cover slip pressure, the sclerites of Cichlidogyrus philander Douëllou, 1993 (Monogenea, Ancyrocephalidae), collected from Pseudocrenilabrus philander (Weber, 1897), were redescribed using SEM. Parasites were collected from Padda Dam, Gauteng, South Africa and soft tissue was digested to release the sclerotized structures. The digested tissue also provided sufficient genetic material for molecular characterization of this species. Cichlidogyrus philander is characterised by a penis with a sharp, curved, lateral termination, an accessory piece with a hook-like extremity that may appear forked terminally, and lack of a visible vagina. The transverse bars have concave and convex surfaces with ribs on the concave surface. The dorsal bar bears fenestrations at the base of the auricles and the ventral and dorsal gripi are dissimilar. Furthermore, the large first pair of uncinuli shows lateral wings on the left side of the base. On top of this wing, a ball-like structure with a small fenestration is visible. Genetic characters derived from the 28S rDNA, the COI mitochondrial DNA and ITS1 rDNA regions distinguish C. philander from all other Cichlidogyrus sequenced species.