Patient quality of life and pain improve after autologous islet transplantation (AIT) for treatment of chronic pancreatitis: 53 patient series at the University of Arizona.

BACKGROUND/OBJECTIVES: Pancreatectomy with autologous islet transplantation has slowly been proving to be an effective way of treating chronic pancreatitis while lessening the effects of the concomitant surgical diabetes of pancreatectomy alone. Assessing patient quality of life and pain after the procedure is particularly important as intractable pain is the main complaint for which patients undergo total pancreatectomy. METHODS: We used the Rand SF-36 and McGill pain questionnaires, and Visual Analogue Scale to assess patients preoperatively for quality of life and pain resulting from life with chronic pancreatitis. After undergoing total pancreatectomy with autologous islet transplantation (TPAIT), patients were followed with surveys administered at 1 month, 6 months, and 1 year to evaluate changes in their quality of life and pain experienced. RESULTS: Significant improvement was reported in all components of every questionnaire within a year after surgery. Furthermore, patient reported mean scores on quality of life were found to fall within the range of the general population. CONCLUSIONS: From our experience with 53 patients at the University of Arizona, after pancreatectomy with autologous islet transplantation patients reported a higher quality of life when compared to preoperative values, as well as reduced levels of pain.

Conservation of a chromosome arm in two distantly related marsupial species.

Marsupials, which diverged from eutherian mammals 150 million years ago (MYA), occupy a phylogenetic position that is very valuable in genome comparisons of mammal and other vertebrate species. Within the marsupials, the Australian and American clades (represented by the tammar wallaby Macropus eugenii, and the opossum Monodelphis domestica) diverged about 70 MYA. G-banding and chromosome painting suggest that tammar wallaby chromosome 6q has homology to opossum chromosome 7q. We tested this conservation by physically mapping the tammar wallaby orthologs of opossum chromosome 7q genes. We isolated 28 tammar wallaby BAC clones that contained orthologs of 16 opossum chromosome 7q genes. We used fluorescence in situ hybridization (FISH) to show that they all mapped specifically to the tammar wallaby chromosome 6q in nearly the same order as their orthologs on opossum chromosome 7q. Thus this chromosome arm is genetically, as well as cytologically, conserved over the 55-80 million years that separate kangaroos and the opossum.

Ischaemic rest pain of the head: a case report.

A 54-year-old man presented with a 3-year history of rest pain of an ischaemic scalp ulcer. Angiography demonstrated that the only blood supply to his head was the left internal carotid artery. Stenting the left subclavian artery and subsequently allowing flow into his left vertebral artery alleviated his symptoms.

The effect of body temperature in a rat model of renal ischemia-reperfusion injury.

BACKGROUND: Renal ischemia-reperfusion injury (IRI) is an unavoidable event in renal transplantation; the effects of IRI can be seen in both the acute and long-term function of the transplanted organ. For this reason, research into the pathophysiology of ischemia-reperfusion is at the forefront of transplantation research. Animal models, particularly in the rat, provide a useful research tool in studying the intricacies of IRI and in evaluating new treatments. We describe a model of right nephrectomy and left renal clamping for 45 minutes and demonstrate the effects of temperature variation during the ischemic period. METHODS: Male Sprague-Dawley rats (under isoflurane anesthesia) underwent bilateral flank incision with removal of the right kidney and clamping of the left renal hilum for 45 minutes. The animals were divided into 3 groups (n=6): group 1 had the procedure performed on a heating mat with no temperature control facilities, group 2 used no heating mat, and group 3 used a rectal temperature-controlled homeothermic blanket system (Harvard Medical, United Kingdom). Temperature was taken every 5 minutes throughout the procedure and blood samples were taken on a daily postoperative basis via tail vein venepuncture. RESULTS: The average temperature at the end of the procedure in group 1 was 39.67 degrees C and the creatinine level at day 3 was 574+/-17.84, in group 2 the temperature was 32.6 degrees C and the creatinine level was 115+/-4.06, and in group 3 the temperature was maintained between 36.5 degrees C-37 degrees C and the serum creatinine level was 329+/-19.18. The temperature of the animal during the ischemia phase of IRI significantly affects the severity of injury. Relative hyperthermia resulted in more severe renal injury and unrecoverable acute renal failure, no source of heat led to a relative hypothermia, and reduction of renal injury. Use of the homeothermic heating blanket led to an increase in creatinine level by day 3, indicating a significant ischemic stimulus; however, by day 10 the creatinine level had returned to normal. CONCLUSION: This illustrates the importance of temperature as a variable in animal models of IRI and thus should be clearly stated in all experimental methods to ensure an appropriate ischemic stimulus and for adequate comparisons between various therapeutic interventions.

FTY720 reduces extracellular matrix expansion associated with ischemia-reperfusion induced injury.

BACKGROUND: Ischemia-reperfusion (IR) is one of the strongest nonimmune factors associated with the development of chronic allograft nephropathy (CAN). This effect is often exacerbated by immunosuppressive medications, most notably cyclosporine. Although traditionally the macrophage was thought to stimulate fibroblast activity in CAN, recent evidence supports a role for lymphocytes. FTY720 is a new immunosuppressant that promotes lymphocyte sequestration into lymph nodes and Peyer's patches. This study investigated the effect of FTY720 on renal fibrosis in the rat following an IR insult (IRI). METHODS: A rat model of IRI was used in which male Sprague-Dawley rats (under isoflurane anaesthesia) underwent bilateral flank incision with removal of the right kidney and clamping of the left renal hilum for 45 minutes. Five groups of animals were studied (n=4): nephrectomy only, IRI only, IRI+FTY720 (1 mg/kg/d), IRI+cyclosporine (15 mg/kg/d), and IRI+FTY 720 (1 mg/kg/d) and cyclosporine (15 mg/kg/d). Animals were humanely killed at 30 days. RESULTS: Serum creatinine (SCr) level was significantly reduced in the FTY720-treated animals. IRI alone produced a significant increase in SCr level compared with neprectomized animals (138 micromol/L vs 55 micromol/L; P<.05). This effect was potentiated by treatment with cyclosporine (173 micromol/L vs 55 micromol/L; P<.05). Treatment with FTY720 significantly reduced SCr level in rats following IRI alone (81 micromol/L vs 138 micromol/L; P<.01) and in rats following IRI + cyclosporine (98 micromol/L vs 173 micromol/L; P<.014). Parallel changes were seen in the levels of proteinuria. Fibrosis was assessed using Masson's trichrome (MT) staining. IRI alone produced a significant increase in MT staining compared with nephrectomized animals (0.92 vs 0.03; P<.05). This effect was potentiated by treatment with cyclosporine (1.12 vs 0.92; P=.022). Treatment with FTY720 reduced the level of MT staining in rats following IRI alone (0.34 vs 0.92; P<.05) and in rats following IRI+cyclosporine (70.34 vs 1.12; P<.05). Levels of TGF-beta1 were considerably reduced in FTY720-treated animals (compared with cyclosporine+IRI and IRI only), either alone (196+/-31 pg/mL vs 1105+/-59 pg/mL and 611+/-38; P<.05) or in conjunction with cyclosporine (423+/-26 pg/mL vs 1105+/-59 pg/mL and 611+/-38; P<.05). CONCLUSION: Our study shows that treatment with FTY720 can reduce renal fibrosis as a result of IRI, both alone and in conjunction with cyclosporine. This provides promising evidence for using FTY720 in a calcineurin-free or reduced-dose immunosuppression protocol in an effort to reduce the incidence of CAN.

Mapping platypus SOX genes; autosomal location of SOX9 excludes it from sex determining role.

In the absence of an SRY orthologue the platypus sex determining gene is unknown, so genes in the human testis determining pathway are of particular interest as candidates. SOX9 is an attractive choice because SOX9 deletions cause male-to-female sex reversal in humans and mice, and SOX9 duplications cause female-to-male sex reversal. We have localized platypus SOX9, as well as the related SOX10, to platypus chromosomes 15 and 10, respectively, the first assignments to these platypus chromosomes, and the first comparative mapping markers from human chromosomes 17 and 22. The autosomal localization of platypus SOX9 in this study contradicts the hypothesis that SOX9 acts as the sex determining switch in platypus.

Sex determination in platypus and echidna: autosomal location of SOX3 confirms the absence of SRY from monotremes.

In eutherian ('placental') mammals, sex is determined by the presence or absence of the Y chromosome-borne gene SRY, which triggers testis determination. Marsupials also have a Y-borne SRY gene, implying that this mechanism is ancestral to therians, the SRY gene having diverged from its X-borne homologue SOX3 at least 180 million years ago. The rare exceptions have clearly lost and replaced the SRY mechanism recently. Other vertebrate classes have a variety of sex-determining mechanisms, but none shares the therian SRY-driven XX female:XY male system. In monotreme mammals (platypus and echidna), which branched from the therian lineage 210 million years ago, no orthologue of SRY has been found. In this study we show that its partner SOX3 is autosomal in platypus and echidna, mapping among human X chromosome orthologues to platypus chromosome 6, and to the homologous chromosome 16 in echidna. The autosomal localization of SOX3 in monotreme mammals, as well as non-mammal vertebrates, implies that SRY is absent in Prototheria and evolved later in the therian lineage 210-180 million years ago. Sex determination in platypus and echidna must therefore depend on another male-determining gene(s) on the Y chromosomes, or on the different dosage of a gene(s) on the X chromosomes.

Vacuum-assisted closure (VAC) therapy in the management of wound infection following renal transplantation.

OBJECTIVES: Wound infection in the setting of immunosuppressed state such as renal transplantation (RT) causes significant morbidity from sepsis, prolongs hospital stay and is expensive. Vacuum-assisted closure (VAC) therapy is a new technique of management of wound based on the principle of application of controlled negative pressure. The aim of this study was to assess the efficacy of VAC therapy in the management of wound infection following RT. MATERIALS AND METHODS: This is a prospective study of a cohort of 180 consecutive RTs performed over a period of 4 years, where the data were retrieved from a prospectively maintained computerised database and case-notes. RESULTS: 9 of 180 (5%) patients developed wound infection following RT which led to cavitations and dehiscence with copious discharge, and refused to heal with conventional treatment. All 9 cases were treated with VAC therapy. The VAC system was removed after a median of 9 (range 3-30) days when discharge from the wound ceased. Four patients were discharged home with portable VAC device and managed on an outpatient basis, where the system was removed after a median 5.5 (range 3-7) days. The median hospital stay after initiation of VAC therapy was significantly shorter (5, range 2-12 days) than on conventional treatment prior to VAC therapy (11, range, 5-20 days) (p=0.003). Complete healing was achieved in all cases. CONCLUSIONS: The use of VAC therapy is an effective and safe adjunct to conventional and established treatment modalities for the management of wound infection and dehiscence following RT. Key words: Renal transplantation, wound infection, vacuum-assisted closure therapy.

Colitis in a renal transplant patient with human herpesvirus-6 infection.

A male patient developed colitis and a thrombotic microangiopathy 3 weeks after renal transplantation. Immunosuppression at the time of presentation was with sirolimus, mycophenolate mofetil, and prednisolone, but without a calcineurin inhibitor. Cytomegalovirus infection was excluded. However, human herpesvirus-6 DNA was detected at high copy number in both blood and colonic epithelium. The patient recovered after reduction in immunosuppression, with nutritional support and ganciclovir therapy.

Construction of a highly enriched marsupial Y chromosome-specific BAC sub-library using isolated Y chromosomes.

The Y chromosome is perhaps the most interesting element of the mammalian genome but comparative analysis of the Y chromosome has been impeded by the difficulty of assembling a shotgun sequence of the Y. BAC-based sequencing has been successful for the human and chimpanzee Y but is difficult to do efficiently for an atypical mammalian model species (Skaletsky et al. 2003, Kuroki et al. 2006). We show how Y-specific sub-libraries can be efficiently constructed using DNA amplified from microdissected or flow-sorted Y chromosomes. A Bacterial Artificial Chromosome (BAC) library was constructed from the model marsupial, the tammar wallaby (Macropus eugenii). We screened this library for Y chromosome-derived BAC clones using DNA from both a microdissected Y chromosome and a flow-sorted Y chromosome in order to create a Y chromosome-specific sub-library. We expected that the tammar wallaby Y chromosome should detect approximately 100 clones from the 2.2 times redundant library. The microdissected Y DNA detected 85 clones, 82% of which mapped to the Y chromosome and the flow-sorted Y DNA detected 71 clones, 48% of which mapped to the Y chromosome. Overall, this represented a approximately 330-fold enrichment for Y chromosome clones. This presents an ideal method for the creation of highly enriched chromosome-specific sub-libraries suitable for BAC-based sequencing of the Y chromosome of any mammalian species.

Spontaneous fulminant gas gangrene.

Gas gangrene is a rare condition, usually associated with contaminated traumatic injuries. It carries a high rate of mortality and morbidity. A number of studies have implicated non-traumatic gas gangrene and colonic neoplasia. This paper reports a patient who presented spontaneously with Clostridium septicum gas gangrene and an occult caecal carcinoma.

Validation of direct intraabdominal pressure measurement using a continuous indwelling compartment pressure monitor.

BACKGROUND: According to recommendations, intraabdominal pressure should be monitored every 8 hours for patients at high risk of abdominal compartment syndrome. Continuous intraabdominal pressure monitoring may be valuable for these patients. METHODS: For 15 patients undergoing laparoscopic surgery, a pressure monitor was introduced after formation of pneumoperitoneum. During the procedure, the laparoscopic insufflator pressure was varied. The pressure monitor values and the time to equilibrium were recorded. RESULTS: Altogether, 152 pressure recordings were taken for the patients studied. The measurements from the insufflator and pressure monitor were compared using a Bland-Altman plot. The mean difference between the techniques was 0.04 +/- 3.8, and 95% of the points from the pressure monitor were within two standard deviations of the mean difference. Pressure changes were essentially "real time." CONCLUSIONS: The intracompartmental pressure monitor provides accurate, rapid, and direct measurement of intraabdominal pressure, and may be a useful tool for continuous intraabdominal pressure measurement among patients at risk of abdominal compartment syndrome.

Cloning and mapping of platypus SOX2 and SOX14: insights into SOX group B evolution.

Group B SOX genes, the closest relatives to the sex-determining gene SRY, are thought to have evolved from a single ancestral SOX B by a series of duplications and translocations. The two SOX B genes SOX2 and SOX14 co-localize to chromosome 3q in humans. SOX2 and SOX14 homologues were cloned and characterized in the platypus, a monotreme mammal distantly related to man. The two genes were found to co-localize to chromosome 1q in this species. Proximity of the two related genes has therefore been conserved for 170 Myr, since humans and platypus diverged. The sequence similarity and conserved synteny of these group B genes provide clues to their origin. A simple model of SOX group B gene evolution is proposed.

The X--a sexy chromosome.

There is new and convincing evidence that the mammalian X chromosome, as well as the Y chromosome, contains an atypically high proportion of genes involved in sex and reproduction (SRR genes). Here we consider alternative explanations for this concentration. One possibility is that a particularly well-endowed autosome was "chosen" for a career as a sex chromosome. Alternatively, the high concentration of SRR genes may have resulted from the accumulation of these genes on the X after the degradation of the Y, either by transposition of autosomal SRR genes to a "selfish X", or by acquisition of SRR functions by widely expressed genes on the X. We suggest experiments to distinguish these possibilities, and speculate on the implications of gathering evidence that genes with other functions, too, are not distributed uniformly over the genome.

Expression and conservation of processed copies of the RBMX gene.

RBMX and RBMY are members of an ancient pair of genes located on the sex chromosomes that encode RNA-binding proteins involved in splicing. These genes have differentiated and evolved separately on the X and Y Chromosomes. RBMY has acquired a testis-specific function, whereas, as shown here, RBMX is ubiquitously expressed and is subject to X inactivation. We have also found that multiple processed copies of RBMX are present in the human genome. RBMX-like sequences (RBMXLs) located on human Chrs 1, 4, 6, 9 (9p13 and 9p24), 11, 20, and X lack introns and thus probably result from retroposition events. We found RBMXLs to be conserved in primates and great apes at corresponding chromosomal locations, indicating that they arose prior to the divergence of human. Some of the RBMXLs show insertions, deletions, and stop codons, which would probably result in nonfunctional proteins. The RBMXL on Chr 20 is deleted in some individuals. Two of the largely intact RBMXLs, located on Chrs 1 and 9p13, are expressed in different tissues and may encode novel proteins involved in splicing in a tissue-specific manner. The RBMXL located at 9p13 is specifically expressed in testis, and to a lesser extent in brain, and may therefore play a role in testis function. This autosomal, testis-specific copy of RBMX could potentially compensate for RBMX that is presumably inactivated in male germ cells, in a manner analogous to autosomal retroposed copies of other X-linked genes.

The human Y chromosome derives largely from a single autosomal region added to the sex chromosomes 80-130 million years ago.

Mapping of human X-borne genes in distantly related mammals has defined a conserved region shared by the X chromosome in all three extant mammalian groups, plus a region that was recently added to the eutherian X but is still autosomal in marsupials and monotremes. Using comparative mapping of human Y-borne genes, we now directly show that the eutherian Y is also composed of a conserved and an added region which contains most of the ubiquitously expressed Y-borne genes. Little of the ancient conserved region remains, and the human Y chromosome is largely derived from the added region.

Ruptured abdominal aortic aneurysm: a novel method of outcome prediction using neural network technology.

BACKGROUND: reported survival following emergency surgery for ruptured abdominal aortic aneurysm (RAAA) varies widely between institutions. This is largely attributable to differences in case mix. The aim of this study was to identify and evaluate a set of prognostic variables that would accurately predict outcome for individual patients from perioperative indices. METHODS: perioperative factors associated with subsequent mortality at our institution were identified by retrospective review of 102 consecutive operations for RAAA over a 7-year period (January 1990 to January 1997). Logistic regression analysis was used to select the most significant variables associated with subsequent mortality. These were used to construct, train, and validate a neural network designed to predict survival from surgery in individual cases on a prospective basis. RESULTS: the 30-day mortality rate was 53%. Multivariate analysis identified four highly significant independent predictors of mortality; preoperative hypotension, intraperitoneal rupture, preoperative coagulopathy, and preoperative cardiac arrest. Using these inputs, the neural network correctly predicted outcome in 82.5% of individual cases. CONCLUSION: a neural network based on just four perioperative variables can accurately predict outcome of RAAA. Prognostic variables should be reported in studies as a measure of the effect of case mix on survival data. Neural networks have potential to aid decision-making relating to outcome for individual cases.