Role of intestine in postsurgical complications: involvement of free radicals

Surgery at any location in the body leads to surgical stress response and alterations in normal body homeostasis. The intestine is extremely sensitive to surgical stress even at remote locations and the gastrointestinal tract plays an important role in the development of postsurgical complications such as sepsis, the systemic immune response syndrome (SIRS), and multiple organ failure syndrome (MOFS). The generation of free radicals and subsequent biochemical alterations at the cellular and subcellular level in the intestine has been suggested to play an important role in this process. These oxidative stress-induced events in the mucosa might act as an initiator of distant organ damage and also facilitate bacterial adherence onto the epithelium and translocation into the systemic circulation. This review attempts to highlight the important role of intestine and oxygen free radicals in initiating post-surgical complications.     

Microtuble organization in Xenopus eggs during the first cleavage and its role in cytokinesis

It has been suggested that the organization of microtubules during mitosis plays an important role in cytokinesis in animal cells. We studied the organization of microtubules during the first cleavage and its role in cytokinesis of Xenopus eggs. First, we examined the immunofluorescent localization of microtubules in Xenopus eggs at various stages during the first cleavage. The astral microtubules that extend from each of the two centrosomes towards the division plane meet and connect with each other at the division plane as cytokinesis proceeds. The microtubular connection thus advances from the animal pole to the vegetal pole, and its leading edge is located approximately beneath the leading edge of the cleavage furrow. Furthermore, an experiment using nocodazole suggests that microtubules have an essential role in advancement of the cleavage furrow, but neither in contraction nor maintenance of the already formed contractile ring which underlies the cleavage furrow membrane. These results suggest that the astral microtubules play an important role in controlling the formation of the contractile ring in Xenopus eggs.     

Natural selection promotes divergence of transferrin among salmonid species

Transferrin is an iron-binding protein that plays an important role in iron metabolism and resistance to bacterial infection in a variety of organisms. A comparison of transferrin coding sequences from four salmonid species shows that the rate of evolution at nonsynonymous sites is significantly higher than the rate at synonymous sites, suggesting that positive natural selection for new alleles has played an important role in the evolution of transferrin in some salmon species. We hypothesize that the selective agent driving rapid divergence is interactions between host transferrin and the iron-scavenging proteins of pathogenic bacteria.     

Increased number of caveolae and caveolin-3 overexpression in Duchenne muscular dystrophy.

Caveolae are small pockets or invaginations localized at the plasma membrane. Caveolins are the principal protein components of caveolae and play an important structural role in the formation of caveolae membranes. Here, we studied by freeze fracture and immunological techniques the spatial organization of caveolae at the muscle cell plasma membrane and the expression of caveolin-3 in Duchenne muscular dystrophy (DMD) muscle fibers. In DMD muscle, we found an increased number of caveolae at the sarcolemma that corresponds to an overexpression of caveolin-3 by immunohistochemistry and by Western blot analysis. These findings suggest a possible role for caveolae and caveolin-3 in the pathogenesis of DMD.     

Nitric oxide participates in the immune response against Neisseria meningitidis serogroup B

The present report explores the role of nitric oxide into the immune response against Neisseria meningitidis serogroup B. Here we show that NO mediates the alphaTNF increase induced by N. meningitidis derived lipopolysaccharides (LPS), at the same time that participates in the bactericidal activity of resting or gammaIFN activated macrophages and plays a role in the specific DTH and IgG response induced by a commercial anti-meningococcal vaccine. Our findings suggest a positive role for NO at the final effector mechanisms and in the early events driving the immunity against N. meningitidis, suggesting also an insight into its role in endotoxic shock.     

A mathematical model for progression and heterogeneity in colorectal cancer dynamics

This paper deals with the development of a mathematical model that describes cancer dynamics at the cellular scale.The selected case study concerns colon and rectum cancer, which originates in colorectal crypts. Cells inside the crypts are assumed to be organized according to a compartmental-like arrangement and to be homogeneously mixing. A mathematical model for cancer progression is proposed here. This model describes the generation of multiple clonal sub-populations of cells at different progression stages in a single crypt.Asymptotic analysis and simulations are developed with an exploratory aim. The obtained results offer some insights into the role played by mutation, proliferation and differentiation phenomena on cancer dynamics. In particular, the acquisition of an additional growing power and a reduction for cellular differentiation seem more likely to be the driving force behind carcinogenesis rather than an increase in the mutation rate. The mutation rate instead seems to affect progression dynamics and intra-tumor heterogeneity. The role played by cells, at different differentiation stages, in the onset and progression of colorectal cancer is highlighted. The results support the fact that stem cells play a key role in cancer development and the idea that transit-amplifying cells could also take on an active role in carcinogenesis.     

Combining native instant photography and photo‐elicitation

The purpose of this study was to explore a combination of two visual strategies to arrive at an emic view of what it is like to be a female re‐entry college student. Both native instant photography and photo‐elicitation were necessary to arrive at the conclusion that re‐entry women have three different types of temporal rhythms in their lives which in turn lead to role overload and guilt. Entering college after having spent many years taking care of a spouse and children creates a role conflict which must be resolved if a woman is to be a successful student. The visual strategies provided an intimate view of this role conflict. Suggestions are made for employing a similar approach to study individuals who are experiencing other types of transition or role conflict.     

Calpain 2 activity increases at the time of implantation in rat uterine luminal epithelial cells and administration of calpain inhibitor significantly reduces implantation sites

The present study investigated the role of calpain 2 in rat uterine luminal epithelial cells during early pregnancy. Calpain 2 is an intracellular calcium-dependent proteolytic enzyme which cleaves numerous focal adhesion proteins. Calpain 2 was concentrated along the basal cell surface of uterine luminal epithelial cells at the predicted site of focal adhesions on day 1 of pregnancy and remained unchanged at the time of implantation as observed by immunofluorescence microscopy. However, Western blotting analysis showed a marked increase in the active form and a significant decrease in the latent form of calpain 2 at the time of implantation. The increase in calpain 2 activity coincides with the disassembly of focal adhesion proteins, talin, paxillin, integrin β1 and β3 from the site of focal adhesions. Intraperitoneal injection of calpain inhibitor, calpain inhibitor l (ALLN), significantly reduced the number of implantation sites, implying that calpain 2 plays an important role in implantation. The present study suggests a role for calpain 2 in the disassembly of focal adhesions, which has been previously shown to play a key role in uterine receptivity for implantation.     

Role modeling in medical education.

Role models play an important part in determining how medical trainees mature professionally. Demonstrating clinical skills at the bedside is the most distinctive characteristic of an effective role model. We discuss how role modeling affects professional identity and career choice and offer several suggestions for improving medical education, including the need for leaders to change the educational climate and culture. If implemented, these changes would enhance our ability to provide medical students with positive role models.     

Beyond KDEL: the role of positions 5 and 6 in determining ER localization

The C-terminal amino acid sequence of a protein plays an important role in determining the endoplasmic reticulum (ER) localization of many soluble proteins that enter the secretory pathway. While it is known that the four amino acids found at the extreme C-terminus of the protein (e.g., KDEL) play a critical role in the interaction with the receptors that mediate retrograde transport back to the ER, other factors within the protein are less well known. Here we show that positions − 5 and − 6 play an important role in determining the ER localization of soluble proteins, with the amino acids at these positions playing an essential role in ER localization of the human protein disulfide isomerase family member, ERp18. Three other naturally occurring C-terminal motifs were also found that work efficiently in ER localization as six-amino-acid variants, but inefficiently as the four-amino-acid variant. Using bimolecular fluorescence complementation, we demonstrate that positions − 5 and − 6 from the C-terminus of the protein play an important role in the recognition of KDEL-like ER retrieval motifs, with the three different human KDEL receptors showing different specificities for changes at these positions for both inefficient and efficient ER localization four-amino-acid motifs.     

Autophagy and plant innate immunity

Plant innate immunity is often associated with specialized programmed cell death at or near the site of pathogen infection. Despite the isolation of several lesion mimic mutants, the molecular mechanisms that regulate cell death during an immune response remain obscure. Recently, autophagy, an evolutionarily conserved process of bulk protein and organelle turnover, was shown to play an important role in limiting cell death initiated during plant innate immune responses. Consistent with its role in plants, several studies in animals also demonstrate that the autophagic machinery is involved in innate as well as adaptive immunities. Here, we review the role of autophagy in plant innate immunity. Because autophagy is observed in healthy and dying plant cells, we will also examine whether autophagy plays a protective or a destructive role during an immune response.     

Role of the hydrophobic moiety of tumor promoters. Synthesis and activity of 2-alkylated benzolactams

The size and position of a hydrophobic moiety on a benzolactam skeleton, which reproduces the active conformation and biological activity of teleocidins, play an important role in the appearance of the activity. Compounds with alkyl groups of various sizes and shapes at the 2-position of benzolactam were synthesized. Structure-activity results indicate that a hydrophobic substituent at the C-2 position plays a critical role in the appearance of biological activities, as in the case of substitution at C-9.     

血管活性肠肽的免疫调节作用

血管活性肠肽作为神经和内分泌系统中一种多功能的神经递质和神经调节因子,在上述两个生理系统中发挥重要的调节作用;同时也对机体免疫系统起着重要的作用,尤其是在局部黏膜免疫中起着一定的调节作用。血管活性肠肽通过它的两个受体VPAC1和VPAC2发挥生物效应。     

The role of Pro-239 in the catalysis and heat stability of subtilisin E

Site-directed mutagenesis was employed to analyze the role of an alpha-helix containing catalytic Ser-221 of subtilisin E. Pro-239 located at the carboxy-terminal end of the alpha-helix was first replaced with Gly to examine the role of Pro-239 in the catalysis and stability of subtilisin E. The mutation was found to decrease both the catalytic rate (kcat) and the heat stability. This result strongly suggests that Pro-239 plays an important role in the maintenance of the alpha-helix, affecting the functioning of the active site. Various amino acid substitutions at position 239 were attempted to obtain the active subtilisins from Gly-239 subtilisin. Lys- and Arg-substitutions were found to result in more active and stable subtilisins than the Gly-239 subtilisin. In particular, the Arg-239 mutant showed enhanced heat stability compared with the wild type. These results demonstrate the important role of the alpha-helix containing catalytic Ser-221 in the catalysis as well as in the heat stability of subtilisin.     

Deletion of the aryl hydrocarbon receptor-associated protein 9 leads to cardiac malformation and embryonic lethality

The aryl hydrocarbon receptor-associated protein 9, ARA9 (also known as XAP2 or AIP1), is a chaperone that is found in complexes with certain xenobiotic receptors, such as the aryl hydrocarbon receptor (AHR) and the peroxisome proliferator-activated receptor alpha (PPARalpha). In an effort to better understand the physiological role of ARA9 outside of its role in xenobiotic signal transduction, we generated a null allele at the Ara9 locus in mice. Mice with a homozygous deletion of this gene die at various time points throughout embryonic development. Embryonic lethality is accompanied by decreased blood flow to head and limbs, as well as a range of heart deformations, including double outlet right ventricle, ventricular-septal defects, and pericardial edema. The early cardiovascular defects observed in Ara9-null mice suggest an essential role for the ARA9 protein in cardiac development. The observation that the developmental aberrations in Ara9-null mice are distinct from those observed for disrupted alleles at Ahr or Pparalpha indicates that the role of ARA9 in cardiac development is independent of its interactions with its known xenobiotic receptor partners.     

Role of the calcium ion and the disulfide bond in the Burkholderia glumae lipase

The role of the Ca²⁺ ion that is present in the structure of Burkholderia glumae lipase was investigated. Previously, we demonstrated that the denatured lipase could be refolded in vitro into an active enzyme in the absence of calcium. Thus, an essential role for the ion in catalytic activity or in protein folding can be excluded. Therefore, a possible role of the Ca²⁺ ion in stabilizing the enzyme was considered. Chelation of the Ca²⁺ ion by EDTA severely reduced the enzyme activity and increased its protease sensitivity, however, only at elevated temperatures. Furthermore, EDTA induced unfolding of the lipase in the presence of urea. From these results, it appeared that the Ca²⁺ ion in B. glumae lipase fulfils a structural role by stabilizing the enzyme under denaturing conditions. In contrast, calcium appears to play an additional role in the Pseudomonas aeruginosa lipase, since, unlike B. glumae lipase, in vitro refolding of this enzyme was strictly dependent on calcium. Besides the role of the Ca²⁺ ion, also the role of the disulfide bond in B. glumae lipase was studied. Incubation of the native enzyme with dithiothreitol reduced the enzyme activity and increased its protease sensitivity at elevated temperatures. Therefore, the disulfide bond, like calcium, appears to stabilize the enzyme under detrimental conditions.     

Promoter recognition by Escherichia coli RNA polymerase. Role of the spacer DNA in functional complex formation

The available evidence suggests that during the process of formation of a functional or "open" complex at a promoter, Escherichia coli RNA polymerase transiently realigns the two contacted regions of the promoter, thus stressing the intervening spacer DNA. We tested the possibility that this process plays an active role in the formation of an open complex. Two series of promoters were examined: one with spacer DNAs of 15 to 19 base-pairs and a derivative for which the promoters additionally contained a one-base gap in the spacer, so as to relieve any stress imposed on the DNA. Consistent with an active role for the stressed DNA in driving open complex formation, we have found that for promoters with a 17-base-pair spacer, the presence of a gap leads to a delay in the formation of an open complex, at a step subsequent to the initial binding of RNA polymerase to the promoter. The results with the other gapped promoters rule out direct binding of RNA polymerase to the region of the gap and indicate an increased flexibility in the gapped DNA. As not all observations with the spacer length series of gapped and ungapped promoters can be interpreted in terms of an active role of the spacer DNA without additional assumptions, such a role must still be considered tentative.