Rotational diffusion of spherical colloids close to a wall.

There is currently no experimental technique available to probe spatially resolved rotational diffusion of nanoparticles in the vicinity of a wall. We present the first experimental study of rotational diffusion of small spherical colloids, using dynamic evanescent wave scattering. A setup is used where the wave vector components parallel and perpendicular to the wall can be varied independently, and an expression is derived for the first cumulant of the intensity correlation function in VH evanescent wave geometry for optically anisotropic spheres. The experimental results are in agreement with theoretical predictions that take particle-wall hydrodynamic interactions into account.

Foliar-Applied Small Molecule that Suppresses Biofilm Formation and Enhances Control of Copper-Resistant Xanthomonas euvesicatoria on Pepper.

We report a small molecule additive, a member of the 2-aminoimidazole (2AI) group that is an analogue of the marine sponge natural product oroidin that suppresses resistance of Xanthomonas euvesicatoria to copper and decreases biofilm formation in an in vitro system. In laboratory experiments, 2AI combined with copper reduced both bacterial multiplication in broth and bacterial recovery on pepper leaf discs of a copper-resistant strain of X. euvesicatoria to a level close to that of a copper-sensitive strain. Compound 2AI used alone exhibited minimal bactericidal activity. In 3 years of field experiments, when combined with a copper-containing material, copper hydroxide (Kocide 3000), and other antibacterial materials, these spray mixtures resulted in decreased bacterial spot foliar disease and increased fruit yields using hybrid bell pepper (Capsicum annuum) cultivars and copper-resistant strains of X. euvesicatoria. This study demonstrates the concept for using small molecules as additives to antibacterial compounds at nonbactericidal concentrations under field conditions that, in the laboratory, were demonstrated to suppress bacterial biofilms and copper-resistant strains.

Aging, yielding, and shear banding in soft colloidal glasses.

Soft colloidal interactions in colloidal glasses are modeled using suspensions of multiarm star polymers. Using a preshearing protocol that ensures a reproducible initial state ("rejuvenation" of the system), we report here the evolution of the flow curve from monotonically increasing to one dominated by a stress plateau, demonstrating a corresponding shear-banded state. Phenomenological understanding is provided through a scalar model that describes the free-energy landscape.

Long-term engraftment following transplantation of pig pancreatic primordia into non-immunosuppressed diabetic rhesus macaques.

BACKGROUND: Transplantation therapy for human diabetes is limited by a shortage of donor organs, and transplant function diminished over time by cell death and limited potential for expansion of beta cells in pancreas or islets. Outcomes are complicated by immunosuppression. A way to overcome supply and expansion problems is to xenotransplant embryonic tissue. Previously, we have shown that beta cells originating from embryonic day (E) 28 (E28) pig pancreatic primordia transplanted into the mesentery of streptozotocin (STZ)-diabetic (type 1) Lewis rats or Zucker Diabetic Fatty (ZDF) diabetic (type 2) rats engraft and normalize glucose tolerance without the need for host immune-suppression. METHODS: In this study, we transplant E28 pig pancreatic primordia in the mesentery of STZ-diabetic rhesus macaques. RESULTS: Long-term engraftment of pig beta cells within liver, pancreas and mesenteric lymph nodes post-transplantation of E28 pig pancreatic primordia into STZ-diabetic rhesus macaques is demonstrated by electron microscopy, positive immune-histochemistry for insulin, and positive RT-PCR and in situ hybridization for porcine proinsulin mRNA. Insulin requirements were reduced in one macaque followed over 22 months post-transplantation and porcine insulin detected in plasma using sequential affinity chromatography, HPLC and mass spectrometry. Of potential importance for application of this transplantation technology to treatment of diabetes in humans and confirmatory of our previous findings in Lewis and ZDF rats, no host immunosuppression is required. CONCLUSIONS: Under selected circumstances, pancreatic primordia elicit a muted immune response relative to more differentiated tissue, such that engraftment occurs in non-immunosuppressed hosts. Our findings that pig pancreatic primordia engraft long-term in non-immunosuppressed STZ-diabetic rhesus macaques establishes the potential for their use in human diabetics.

Gastric emptying in post-thoracotomy patients receiving a thoracic fentanyl-bupivacaine epidural infusion.

BACKGROUND AND OBJECTIVE: The effect of anaesthesia and surgery on gastric emptying is not constant, and this has not been previously studied in patients undergoing thoracotomy for lung resection with a bupivacaine-fentanyl epidural infusion for analgesia. There are important implications in this group of patients with regards to the recommencement of important oral medication as well as the risks of aspiration of gastric contents. The study examined gastric emptying in these patients until the second postoperative day. METHODS: In a prospective repeated measures study, the effect of fentanyl-bupivacaine epidural analgesia at the mid-thoracic level on gastric emptying was assessed in 11 patients undergoing thoracotomy for lung resection. Gastric emptying was measured using a paracetamol absorption technique. Patients acted as their preoperative controls and were assessed 4 h postoperatively and on the second postoperative day. RESULTS: The mean (SEM) maximum plasma paracetamol concentration was 204.6 (20.4) micromol L(-1) before operation, 61 (9.5) micromol L(-1) 4 h postoperatively and 114.3 (22.6) micromol L(-1) on the second postoperative day. Mean (SEM) paracetamol absorption at 120 min was 15,638 (1441) micromol min L(-1) preoperatively, 5731 (821) micromol min L(-1) 4 h postoperatively and 9325 (1759) micromol min L(-1) on the second postoperative day. Postoperative values were significantly (P < 0.005) less than the preoperative values. CONCLUSIONS: After thoracotomy, gastric emptying was delayed until at least the second postoperative day in patients receiving mid-thoracic fentanyl-bupivacaine epidural analgesia.

Transplantation of metanephroi after preservation in vitro.

To determine whether transplanted metanephroi grow, differentiate, and function in hosts after preservation in vitro, we implanted metanephroi from embryonic day 15 (E15) Sprague-Dawley rat embryos into the omentum of nonimmunosuppressed uninephrectomized Sprague-Dawley (host) rats. Metanephroi were either implanted directly or suspended in ice-cold University of Wisconsin (UW) preservation solution with or without added growth factors for 3 days before implantation. The size and extent of tissue differentiation preimplantation of E15 metanephroi implanted directly were not distinguishable from the size and differentiation of metanephroi preserved for 3 days. In contrast, E16 metanephroi were larger than E15 metanephroi preserved for 3 days. E16 metanephroi or E13 metanephroi grown in organ culture for 3 days contained more differentiated nephron structures than those in E15 metanephroi preserved for 3 days. By 4 wk posttransplantation, metanephroi that had been preserved for 3 days had grown and differentiated such that glomeruli, proximal and distal tubules, and collecting ducts with normal structure had developed. At 12 wk posttransplantation, inulin clearances of preserved metanephroi were comparable to those of metanephroi that had been implanted directly. Addition of growth factors to the UW solution enhanced inulin clearances. Here we show for the first time that functional kidneys develop from metanephroi transplanted from rat embryos to adult rats after as long as 3 days of preservation in vitro.

Sevoflurance: approaching the ideal inhalational anesthetic. a pharmacologic, pharmacoeconomic, and clinical review.

Sevoflurane is a safe and versatile inhalational anesthetic compared with currently available agents. Sevoflurane is useful in adults and children for both induction and maintenance of anesthesia in inpatient and outpatient surgery. Of all currently used anesthetics, the physical, pharmacodynamic, and pharmacokinetic properties of sevoflurane come closest to that of the ideal anesthetic (200). These characteristics include inherent stability, low flammability, non-pungent odor, lack of irritation to airway passages, low blood:gas solubility allowing rapid induction of and emergence from anesthesia, minimal cardiovascular and respiratory side effects, minimal end-organ effects, minimal effect on cerebral blood flow, low reactivity with other drugs, and a vapor pressure and boiling point that enables delivery using standard vaporization techniques. As a result, sevoflurane has become one of the most widely used agents in its class.

Airway fire due to diathermy during tracheostomy in an intensive care patient.

We describe a case of airway fire in an 83-year-old, critically ill patient. The fire occurred during a surgical tracheostomy under general anaesthesia, following ignition of the tracheal tube by diathermy. After debridement of the burnt tissue and treatment with intravenous antibiotics and glucocorticoids, the patient's respiratory function worsened initially. The patient eventually recovered without long-term sequelae and was discharged from the intensive care unit. The circumstances of this and other similar incidents are reviewed, as are the suggested methods for preventing this frightening occurrence.

Transplantation of rat metanephroi into mice.

To determine whether transplanted metanephroi grow and differentiate after implantation into the omentum in hosts of a different species, we implanted metanephroi from embryonic day 15 (E15) rat embryos into uninephrectomized mice (hosts). Some host mice received human CTLA4Ig (hCTLA4Ig), anti-CD45RB, and anti-CD154 (tolerance-inducing agents). E15 metanephroi contained only metanephric blastema, segments of ureteric bud, and primitive nephrons with no glomeruli. Rat metanephroi did not grow or differentiate in mice that received no tolerance-inducing agents. However, by 2 wk posttransplantation in mice that received hCTLA4Ig, anti-CD45RB, and anti-CD154, metanephroi from E15 rats had enlarged, become vascularized, and formed mature tubules and glomeruli. Rat metanephroi contained cells that stained specifically for mouse CD31, a marker for sprouting endothelial cells. Some rat glomerular capillary loops stained positively for mouse CD31. Here, we show that chimeric kidneys develop from metanephroi transplanted rat-->mouse and that glomeruli are vascularized, at least in part, by host vessels.

The prevalence of religious coping among persons with persistent mental illness.

OBJECTIVE: The purpose of this study was to examine the prevalence of religious coping among persons with persistent mental illness and to gain a preliminary understanding of the relationship between religious coping and symptom severity and overall functioning. METHODS: A total of 406 individuals who were diagnosed as having a mental illness and who were patients at one of 13 Los Angeles County mental health facilities completed a survey consisting of the Religious Coping Index, the Symptom Checklist 90-R (SCL-90), the Global Assessment of Functioning (GAF) scale, and a 48-item demographic questionnaire. RESULTS: More than 80 percent of the participants used religious beliefs or activities to cope with daily difficulties or frustrations. A majority of participants devoted as much as half of their total coping time to religious practices, with prayer being the most frequent activity. Specific religious coping strategies, such as prayer or reading the Bible, were associated with higher SCL-90 scores (indicating more severe symptoms), more reported frustration, and a lower GAF score (indicating greater impairment). The amount of time that participants devoted to religious coping was negatively related to reported levels of frustration and scores on the SCL-90 symptom subscales. CONCLUSIONS: The results of the study suggest that religious activities and beliefs may be particularly compelling for persons who are experiencing more severe symptoms, and increased religious activity may be associated with reduced symptoms. Religion may serve as a pervasive and potentially effective method of coping for persons with mental illness, thus warranting its integration into psychiatric and psychological practice.

Transplantation of metanephroi across the major histocompatibility complex in rats.

To determine whether transplanted metanephroi grow, differentiate, and function in hosts that differ in major histocompatibility complex loci (RT1 loci in rats) from donors in a defined way, we implanted metanephroi from embryonic day (E) 15 PVG (RT1(c)) rat embryos into the omentum of nonimmunosupressed uninephrectomized PVG-RT1(avl) (host) rats. By 4 wk posttransplantation, metanephroi had grown and differentiated such that glomeruli, proximal and distal tubules, and collecting ducts had normal structure and ultrastructure. At 12 wk posttransplantation, weights of metanephroi were 54 +/- 8 mg. Inulin clearances were 0.9 +/- 0.3 microl. min(-1). 100 g rat wt(-1). In vitro, splenocytes from PVG rats stimulated the proliferation of cells originating from both PVG-RT1(avl) rats in which a transplant had been performed and PVG-RT1(avl) rats with no transplant. Full-thickness PVG-RT1(avl) skin engrafted normally on PVG-RT1(avl) rats in which PVG metanephroi had been previously implanted and metanephroi retained a normal appearance. In contrast, skin from PVG rats sloughed, and the tubular architecture of metanephroi was obliterated by a mononuclear cell infiltrate consistent with acute rejection. Here we show for the first time that functional chimeric kidneys develop from metanephroi transplanted across the MHC into nonimmunosupressed hosts and provide evidence that a state of peripheral immune tolerance secondary to T cell "ignorance" permits their survival.

Characterization of cationic vector-based gene delivery vehicles using isothermal titration and differential scanning calorimetry.

Within the past 10 years, major advances in the design and development of differential scanning calorimeters (DSC) (1) and isothermal titration calorimeters (ITC) (2) have resulted in an unparalleled level of sensitivity, stability, and reproducibility in calorimetric measurements of large molecules. These improvements have allowed the thermal stability and ligand binding processes of biological macromolecules to be thermodynamically characterized with speed, accuracy, and convenience. With their increasing commercial availability, experiments that were previously limited to specialist calorimetry laboratories can now be routinely performed by most investigators.

Ty1-copia-like retrotransposons of tomato (Lycopersicon esculentum Mill.).

We have used a PCR and cloning strategy to identify Ty1-copia-like retrotransposons in tomato, Lycopersicon esculentum Mill. Using degenerate oligonucleotide primers corresponding to conserved domains of the Ty1-copia retrotransposon reverse transcriptase (RT), fragments of about 260 bp were obtained by PCR amplification. Sequences of 20 cloned amplification fragments showed similarity to retrotransposon sequences. The copy number for total tomato Ty1-copia-like RT population was estimated to be approximately 2500 and may account for about 1.5% of the tomato genome. Copy numbers for four of the individual RT clones ranged from 20 to 1400 copies. A comparison of the conceptual translations of the RT sequences identified four clusters as well as three sequences which were ungrouped. When compared to RT sequences reported from several other sources, the tomato RT population was found to be widely dispersed with the majority of the RT sequences from Lycopersicon species delineated by the four tomato cluster groups. The gag region of a tomato retrotransposon was cloned from PCRs with primers based on the Tnt1 retrotransposon of tobacco. The tomato clone (pTom1.1) had 81% sequence similarity to the Tntl gag region. Several pTom1.1 sequences are present in other solanaceous species as indicated by Southern hybridization.

Cloning of the Atlantic salmon (Salmo salar) estrogen receptor-alpha gene.

A cDNA clone encoding most of an Atlantic salmon (Salmo salar) estrogen receptor (ER) was obtained from a liver cDNA library and the remainder of the coding sequence from the gene was isolated from a genomic library. Sequence comparisons showed that the cloned gene represents ER-alpha. Expression of the ER-alpha gene in male and female salmon parr was analysed by RT-PCR. Highest expression was found in brain and liver, with lower levels of ER-alpha mRNA present in all other tissues tested. There was little difference in expression of ER-alpha between male and female.

Insulin-like growth factor I regulates renal development in rodents.

Blocking the action of insulin-like growth factor I (IGF I) impairs kidney development in vitro. However, no renal developmental abnormalities have been reported in newborn transgenic mice that do not express IGF I (Igf1-/-) mice. Ninety-five percent of Igf1-/- mice die immediately following birth. Kidney development continues following birth in rodents. To readdress the question of the participation of IGF I in the process of kidney development, we measured nephron numbers in developed kidneys from Igf1-/- mice that survived past birth, and using a second model of kidney development, characterized the effect of IGF I infused into rat hosts on the renal function of transplanted metanephroi. Igf1-/- mice were born with grossly normal kidneys. At 77 +/- 10 days after birth, Igf1-/- mice that survived were approximately 28% the weight of wild-type (WT) littermates and had proportionally smaller kidneys. The number of nephrons per kidney was reduced by approximately 20% in Igf1-/- mice. Glomerular size was also reduced in Igf1-/- mice. In untreated host rats, neither the size nor inulin clearance of transplanted metanephroi changed significantly from 12-28 weeks postimplantation. The administration of IGF I to hosts did not affect the size of transplanted metanephroi measured at 12-16 weeks following implantation. However, inulin clearances were increased significantly by the administration of IGF I to hosts. Our findings 1) indicate that IGF I plays a role in determining nephron number, 2) suggest that it enhances function in developing kidneys, and 3) establish the potential for the pharmacological use of IGF I to enhance the growth and function of transplanted metanephroi.

Transplantation of developing metanephroi into adult rats.

BACKGROUND: Transplantation of developing metanephroi into adult hosts has been proposed as a means to augment host renal function. METHODS: We implanted whole metanephroi from embryonic day 15 (E15) rats subcapsularly in kidneys or into the omentum of non-immunosupressed adult rat hosts. At the time of implantation, some host rats underwent unilateral nephrectomy (UNX) or unilateral nephrectomy and partial contralateral renal infarction (1 1/2 NX). E15 metanephroi contained only metanephric blastema, segments of ureteric bud, and primitive nephrons with no glomeruli. RESULTS: Four to six weeks post-implantation, metanephroi from E15 rats had enlarged, become vascularized, and had formed mature tubules and glomeruli. Ureters of metanephroi transplanted into the omentum were anastomosed to hosts' ureters that remained after UNX. Four weeks following ureteroureterostomy, the contralateral kidney was removed. Inulin clearances of seven metanephroi implanted into UNX hosts averaged 0.11 +/- 0.02 microliters/min/100 g (2.42 +/- 0.70 microliters/min/g kidney wt) and the creatinine clearances averaged 0.65 +/- 0.18 microliters/min/100 g. Metanephroi weighed 71 +/- 15 mg (approximately 4% of the contralateral native kidney). The transplanted metanephroi were vascularized by arteries originating from the omentum. Both weights of transplanted metanephroi (145 +/- 24 mg) and inulin clearances of transplanted metanephroi (30.1 +/- 8.7 microliters/min/g kidney weight) were significantly increased in rats that underwent 1 1/2 NX compared to UNX. In contrast, transplantation of developed kidneys resulted in rejection. CONCLUSIONS: Our findings establish that functional chimeric kidneys develop from metanephroi transplanted in adult hosts.

Cloning and unusual expression profile of the aldolase B gene from Atlantic salmon.

A full-length clone of the aldolase B gene has been isolated from a cDNA library constructed from liver of Atlantic salmon (Salmo salar). Sequencing showed that the clone encodes a typical aldolase B, possessing a number of amino acid residues which are seen in aldolase B, but not in other aldolase isoforms. RT-PCR analysis showed that the gene is expressed in liver, kidney and intestine as expected. However, in contrast to mammalian and avian aldolase B, expression was also found in a number of other tissues. Levels of aldolase B mRNA in liver and kidney were not significantly altered during smoltification, the transformation of freshwater-dwelling salmon (parr) into saltwater-adapted salmon (smolts).

Metanephric osteopontin regulates nephrogenesis in vitro.

Renal expression of osteopontin is enhanced in the setting of acute ischemic injury. Because of the parallels that exist between recovery from renal ischemia and renal development, we characterized the role that osteopontin plays during metanephrogenesis in the rat. Osteopontin mRNA is present in kidneys obtained from rat embryos as early as embryonic day 13 (E13). Immunohistochemical staining of metanephroi obtained from E16 rat embryos and metanephroi obtained from E13 embryos and cultured for 3 days in vitro demonstrated that osteopontin is expressed both in the developing nephron and in the ureteric bud. Addition of anti-osteopontin antibodies to metanephric organ cultures results in failure of the metanephric blastema to undergo normal tubulogenesis. Addition of the arginine-glycine-aspartic acid-containing peptide, cyclo-RGDfV, or the anti-alpha(v)beta3-integrin antibody, LM609, to cultures has a similar effect. These findings establish that osteopontin is produced within the rat metanephros during development in vivo and suggest that the binding of osteopontin to the alpha(v)beta3-integrin is required for tubulogenesis to occur in vitro. Blastemal cells within metanephroi cultured in the presence of OP199 manifest increased apoptosis compared with controls. It is possible that osteopontin plays an important anti-apoptotic role during the process of metanephric blastema condensation that is a prerequisite for the formation of nephrons in vivo.

Two core promotor mutations identified in a hepatitis B virus strain associated with fulminant hepatitis result in enhanced viral replication.

Viral mutations have been implicated in alteration of the biological phenotype of hepatitis B virus (HBV). We recently cloned and sequenced the viral genome of an HBV strain associated with an outbreak of fulminant hepatitis (FH strain). The FH strain contained numerous mutations in all genomic regions and was functionally characterized by a more efficient encapsidation of pregenomic RNA leading to highly enhanced replication. To define the responsible mutation(s) for the enhanced replication, we introduced individual mutations of the FH strain into a wild-type construct by oligonucleotide-directed mutagenesis. Analysis of viral replication showed that two adjacent mutations in the HBV core promotor (C to T at nucleotide 1768 and T to A at nucleotide 1770) led to high level replication. Similar to the FH strain, this mutant displayed the phenotype of enhanced encapsidation of pregenomic RNA. Functional studies in an encapsidation assay demonstrated that the identified mutations resulted in a minor increase of pregenomic RNA transcription (two- to threefold) and a major transcription-independent enhancement (> 10-fold) of viral encapsidation. Our results demonstrate that the two adjacent mutations in the HBV core promotor region are responsible for the enhanced replication of the FH strain. These two mutations, outside the previously described encapsidation signal, core, and polymerase polypeptides, appeared to affect a novel genetic element involved in viral encapsidation.