Association between thyroid dysfunction and dysglycaemia: a prospective cohort study.

AIMS: To compare the incidence of hyperglycaemia among participants with low, elevated and normal serum thyroid-stimulating hormone concentration, as well as the incidence of abnormal thyroid function test results among participants with normal blood glucose and those with hyperglycaemia. METHODS: In a prospective study, a cohort of 72 003 participants with normal, low and elevated serum thyroid-stimulating hormone concentration were followed from the study beginning to the first report of diabetes and prediabetes. A proportional hazards regression model was used to calculate the hazard ratios and 95% CIs for each outcome, adjusting for age, sex, education level, smoking, alcohol consumption and obesity. Analyses for the association between dysglycaemia and incident abnormal thyroid function test were also conducted. RESULTS: During a median 2.6 year follow-up, the incident rates for dysglycaemia, particularly prediabetes, were substantially higher in participants with elevated thyroid-stimulating hormone concentrations at baseline, while the rates for participants with normal and low thyroid-stimulating hormone were similar. After controlling for risk factors, participants with elevated thyroid-stimulating hormone retained a 15% increase in risk of prediabetes (adjusted hazard ratio 1.15, 95% CI 1.04-1.26), but were not at greater risk of diabetes (adjusted hazard ratio 0.96, 95% CI 0.64-1.44). By contrast, participants with normal and low thyroid-stimulating hormone concentrations had similar dysglycaemia risks. Participants with diabetes and prediabetes were not at greater risks of developing abnormal thyroid function test results when compared with participants with euglycaemia. CONCLUSIONS: People with elevated serum thyroid-stimulating hormone concentration are at greater risk of developing prediabetes. Whether this includes a greater risk of developing frank diabetes may require an extended period of follow-up to clarify.

Protection against lethal enterovirus 71 infection in newborn mice by passive immunization with subunit VP1 vaccines and inactivated virus.

Enterovirus 71 (EV71), the newest member of Enteroviridae, is notable for its etiological role in epidemics of severe neurological diseases in children. Developing effective vaccines is considered a top choice among all control measures. We compared the inactivated virus vaccine (10 microg protein/mouse) with subunit vaccines--VP1 DNA vaccine (100 microg/mouse) or recombinant VP1 protein (10 microg/mouse)--in its ability to elicit maternal antibody and to provide protection against lethal infection of EV71 in suckling mice. Prior to gestation, all three groups of vaccinated dams possessed similar levels of neutralizing antibody. With a challenge dose of 2300 LD(50) virus/mouse, suckling mice born to dams immunized with inactivated virus showed 80% survival. The subunit vaccines provided protection only at a lower challenge dosage of 230 LD(50) per mouse, with 40% survival for DNA vaccine and 80% survival for VP1 protein. The cytokine profile produced by splenocytes showed a high level of IL-4 in the inactivated virus group, high levels of IFN-gamma and IL-12 in the DNA vaccine group, and high levels of IL-10 and IFN-gamma in the VP1 protein group. Overall, the inactivated virus elicited a much greater magnitude of immune response than the subunit vaccines, including total IgG, all four IgG subtypes, and T-helper-cell responses; these antibodies were shown to be protective against lethal infection when passively transferred to susceptible newborn mice. Our data indicated that inactivated virus is the choice of vaccine preparation capable of fulfilling the demand for effective control, and that VP1 subunit vaccines remain promising vaccine strategies that require further refinement.

Association of pityriasis rosea with human herpesvirus-6 and human herpesvirus-7 in Taipei.

BACKGROUND AND PURPOSE: Pityriasis rosea (PR) is a common papulosquamous skin disease with unknown etiology. The possible relationship of PR with human herpesvirus infection (HHV) has been extensively studied. This study used the polymerase chain reaction (PCR) to investigate the presence of human herpesvirus 6 and 7 (HHV-6 and HHV-7) in 41 PR patients from two hospitals in Northern Taiwan. The epidemiologic features of PR in patients were also studied. METHODS: A total of 41 PR patients (11 males, 30 females) were enrolled in this study from April 1999 to March 2000. PCR of skin biopsy specimens from 24 PR patients was used to identify the existence of HHV-6 and HHV-7. Viral culture from PR biopsy specimens was also performed. Blood from these patients was sampled for Venereal Disease Research Laboratory tests. Skin biopsies from 20 age- and sex-matched controls with other skin diseases were also subjected to PCR study. RESULTS: The ages of the 41 PR patients ranged from 8 to 62 years. An increased incidence (17/41) of PR episodes was observed during the spring. Both HHV-6 and HHV-7 DNA was below the limit of detection in all biopsy specimens from patients and healthy controls. Viral culture for HHV was negative in all patients. CONCLUSION: The epidemiologic features of PR in this series are comparable to other studies except for an exaggerated female predominance (male:female ratio 1:2.7). Our data indicate a lack of association between HHV-6 and HHV-7 infection and PR.

Expression of capsid [correction of caspid] protein VP1 for use as antigen for the diagnosis of enterovirus 71 infection.

To produce enterovirus 71 antigen for diagnostic purposes, the gene encoding the entire capsid protein VP1 was amplified by reverse transcription-polymerase chain reaction (RT-PCR), cloned and expressed in Escherichia coli as a poly-histidine fusion protein. Western blotting experiments with sera from patients with enterovirus 71 infection indicated that immunoglobulin G (IgG) and IgM antibodies bound to a single polypeptide VP1. According to these results, IgM anti-VP1 appeared in sera of patients with a symptomatic enterovirus 71 acute infection, whereas IgG anti-VP1 was present in sera of past infection. This finding suggests that detecting IgG and IgM immune responses against linear epitopes of recombinant VP1 is an effective means of determining the different phases of enterovirus 71 infection. In addition, sera containing coxsackie virus 16 (CA16) antibodies did not cross-react with the recombinant VP1 of enterovirus 71, despite the homology between VP1 proteins of both viruses. Comparison with reference PCR and neutralization assays showed these antibody tests to be appropriate for the serodiagnosis of enterovirus 71 infection.

Diagnosis of respiratory tract viruses in 24 h by immunofluorescent staining of shell vial cultures containing Madin-Darby Canine Kidney (MDCK) cells.

Nine hundred and seventy-eight clinical specimens were examined taken from patients with respiratory tract viruses (RV)-like syndrome between November 1996 and July 1998. The study was undertaken to evaluate the effectiveness of centrifuge-enhanced shell vial cultures (SVC) containing Madin-Darby Canine Kidney (MDCK) cells, combined with immunofluorescent (IF) staining in 24 h. This technique rapidly detects and identifies respiratory tract viruses. The conventional tube culture system with multiple cell lines would ordinarily detect RV within 3-30 days. The SVC/IF method using single cell line (MDCK cells) allowed detection of 81.5% of influenza A virus, 72% of parainfluenza virus, 82.6% of respiratory syncytial virus (RSV) and 79.6% of adenovirus in 24 h.

Characterization of the inhibition of protein phosphatase-1 by DARPP-32 and inhibitor-2.

Phospho-DARPP-32 (where DARPP-32 is dopamine- and cAMP-regulated phosphoprotein, Mr 32,000), its homolog, phospho-inhibitor-1, and inhibitor-2 are potent inhibitors (IC50 approximately 1 nM) of the catalytic subunit of protein phosphatase-1 (PP1). Our previous studies have indicated that a region encompassing residues 6-11 (RKKIQF) and phospho-Thr-34, of phospho-DARPP-32, interacts with PP1. However, little is known about specific regions of inhibitor-2 that interact with PP1. We have now characterized in detail the interaction of phospho-DARPP-32 and inhibitor-2 with PP1. Mutagenesis studies indicate that within DARPP-32 Phe-11 and Ile-9 play critical roles, with Lys-7 playing a lesser role in inhibition of PP1. Pro-33 and Pro-35 are also important, as is the number of amino acids between residues 7 and 11 and phospho-Thr-34. For inhibitor-2, deletion of amino acids 1-8 (I2-(9-204)) or 100-204 (I2-(1-99)) had little effect on the ability of the mutant proteins to inhibit PP1. Further deletion of residues 9-13 (I2-(14-204)) resulted in a large decrease in inhibitory potency (IC50 approximately 800 nM), whereas further COOH-terminal deletion (I2-(1-84)) caused a moderate decrease in inhibitory potency (IC50 approximately 10 nM). Within residues 9-13 (PIKGI), mutagenesis indicated that Ile-10, Lys-11, and Ile-13 play critical roles. The peptide I2-(6-20) antagonized the inhibition of PP-1 by inhibitor-2 but had no effect on inhibition by phospho-DARPP-32. In contrast, the peptide D32-(6-38) antagonized the inhibition of PP1 by phospho-DARPP-32, inhibitor-2, and I2-(1-120) but not I2-(85-204). These results indicate that distinct amino acid motifs contained within the NH2 termini of phospho-DARPP-32 (KKIQF, where italics indicate important residues) and inhibitor-2 (IKGI) are critical for inhibition of PP1. Moreover, residues 14-84 of inhibitor-2 and residues 6-38 of phospho-DARPP-32 share elements that are important for interaction with PP1.

A new solution for life without blood. Asanguineous low-flow perfusion of a whole-body perfusate during 3 hours of cardiac arrest and profound hypothermia.

BACKGROUND: The benefits of hypothermia for preventing ischemic injury are well known, but its application in surgery to protect the whole body during procedures requiring circulatory arrest is currently limited to < 1 hour at 15 degrees C using 50% hemodilution. In a significant departure from previous methods, we have developed a technique of asanguineous blood substitution with low-flow perfusion and cardiac arrest at < 10 degrees C in a canine model. Our approach has been to design a hypothermic blood substitute that would protect the brain and visceral organs during several hours of bloodless perfusion. Two different solutions have been designed to fulfill separate requirements in the procedure. METHODS AND RESULTS: With the use of extracorporeal cardiac bypass, 14 adult dogs were exsanguinated during cooling; 11 dogs were blood substituted using in combination the "purge" and "maintenance" solutions (group 1), and 3 dogs were perfused throughout with the "purge" solution alone as controls (group 2). After cardiac arrest, the solutions were continuously circulated for 3 1/2 hours by the extracorporeal pump (flow rate, 40 to 85 mL.kg-1.min-1; mean arterial blood pressure, 25 to 40 mm Hg). The temperature was maintained at < 10 degrees C (nadir, 6.6 +/- 0.1 degrees C) for 3 hours, and the hematocrit was kept at < 1% before controlled rewarming and autotransfusion. In the experimental group, the heart always started spontaneously in the temperature range of 11 degrees C to 27 degrees C, and 8 animals have survived long-term (current range, 14 to 110 weeks) without any detectable neurological deficit. In contrast, two control animals survived after extensive and aggressive cardiac resuscitation efforts; after surgery they exhibited transient motor and sensory deficits for approximately 1 week. Evaluation of biochemical and hematological parameters showed only a transient and inconsequential elevation in enzymes (eg, brain, liver, cardiac) in group 1 compared with the markedly greater elevations in group 2. For example, immediate postoperative values (mean +/- SEM) for lactate dehydrogenase were 114 +/- 10 for group 1 versus 490 +/- 210 for group 2 (P < .03); for SGOT, values were 93 +/- 18 for group 1 versus 734 +/- 540 for group 2 (P < .05). On day 1 for creatine kinase (CK), the group 1 value was 7841 +/- 2307 versus 71,550 +/- 2658 for group 2 (P = .03), and for CK-BB, the group 1 value was 108 +/- 22 versus 617 +/- 154 for group 2 (P = .03). Neurological evaluation using deficit scores (NDS) was based on a modification of the Glasgow Coma Scale score: 0, normal; 1, minimal abnormality; 2, weakness; 3, paralysis; 4, coma; and 5, death. At days 1 and 2 after surgery, NDS (mean +/- SEM) were 0 +/- 0 for the experimental group versus 1.5 +/- 0.5 for the control group. At days 3 and 7 after surgery, NDS were 0 +/- 0 for group 1 versus 1.0 +/- 1.0 for group 2. CONCLUSIONS: The faster neurological recovery of dogs treated with the "intracellular-type" maintenance solution supports the biochemical data showing the benefits of this type of blood substitute for extending the safe limits of hypothermic cardiac arrest procedures to > 3 hours.

Molecular cloning of the common carp (Cyprinus carpio) rhodopsin cDNA.

A recombinant phage clone containing a 1584 nucleotides rhodopsin cDNA was screened from a carp retinal cDNA library. The inserted DNA consisting of a single open reading frame of 1062 nucleotides at positions 72 to 1133 encodes a 354 amino acid polypeptide. The deduced amino acid sequence of carp rhodopsin showed 95.7, 85.5 and 74.4% identity with that of goldfish, sand goby and lamprey, respectively. The sites of palmitoylation, glycosylation, disulfide bond formation and Schiff base formation in the putative rhodopsin are all conserved.

A pilot investigation of poloxamer 407 for the prevention of leptomeningeal adhesions in the rabbit.

Leptomeningeal adhesion formation frequently complicates operations and diseases of the central nervous system. Chronic adhesive arachnoiditis may follow intraspinal surgery for disc, tumor, and closure of myelomeningocele, eventually producing pain and declining neurological status of the patient. Reoperation for scar removal is seldom successful as the arachnoidal adhesions reform. Poloxamer 407 (P407) has been shown to reduce postoperative peritoneal adhesion formation in rats and golden hamsters. In a rabbit model, we investigated the potential of P407 to prevent the production of arachnoidal adhesions and nerve root scarring following laminectomy and surgical meningeal injury. The lumbar spinal roots of 8 New Zealand white rabbits were surgically isolated under magnification. One root sleeve axilla was opened and immediately closed with 10-0 suture (control site) and a second root sleeve axilla was opened, P407 injected, and closed with 10-0 suture (treatment site). Five of 7 rabbits treated with P407 and followed for 7-42 days showed no arachnoidal adhesions at the level of the nerve root. Four New Zealand white rabbits had the lamina removed, and the dura over the spinal cord was opened at two sites separated by one to two lumbar segments. At one site P407 was inserted beneath the dura following durotomy, and the other site was opened in a similar fashion and immediately closed without the insertion of P407. There was a 50% reduction in leptomeningeal adhesion formation with the use of P407. P407 may be useful in neurosurgery for the prevention of arachnoidal adhesions.

CO2-laser radiation damage of the arterial wall.

This study describes the effects of CO2 laser radiation on the histology of the normal rabbit arterial wall, using models that simulate laser angioplasty and anastomosis. Rabbit arteries were exposed to laser treatments similar to those used clinically; 40, 0.5 sec pulses of 40-60 mW, CO2 continuous wavelength laser, or a 1/2-circumferential laser anastomosis with a 60-80 mW continuous pulse. Aneurysms developed in 8 of 22 femoral, 1 of 22 carotid, and no controls at 12 week. There were small breaks in the internal elastic lamina with atrophy, loss of muscularis, "packing" of the elastica, thinning of the muscularis at the damage site, and enlargement of the arterial diameter. Aneurysms developed in one femoral and no carotid anastomosed artery. Laser anastomoses demonstrated more muscle damage and loss, with extensive scarring and a wider area of elastic loss than the controls. The intima was reestablished with focal reduplication of the internal elastic lamina. There were no histologic differences between the arteries which developed aneurysms and those which did not in either series. These results suggest that low power laser damage of the arterial wall consists mainly of destruction of the muscularis propria, with minimal damage to the elastica.

Pulmonary venous anastomosis in lung transplantation without donor left atrium. Experimental and clinical results.

Single lung transplantation now is a therapeutic option for some patients with end-stage lung disease. Cyclosporine immunosuppression and refinements in bronchial anastomosis have been responsible for recent successes. Since 1953, the usual pulmonary venous anastomosis, both in experimental animals and in humans, has been an atrium-to-atrium connection. This technique may limit the availability of usable donor lungs, since the donor heart, along with the atrium, is usually harvested for another recipient. Although techniques can be developed to allow both transplant teams to harvest atrial tissue, this study was undertaken to determine if, in fact, anastomosis with donor left atrium is necessary. Twenty-four dogs were anesthetized and a left thoracotomy performed. After heparinization (3 mg/kg), the pulmonary artery and left atrium were occluded. One of four different pulmonary venous anastomoses was performed at 3.5x magnification: superior pulmonary vein end to end (group I), inferior pulmonary vein end to end (group II), superior pulmonary vein implantation into left atrium (group III), and left atrium-to-left atrium anastomosis as control (group IV). Everting mattress sutures of 7-0 polypropylene were used in groups I, II, and III and 6-0 in group IV. Average crossclamp time for group I, group II, and group IV was 20 minutes. The average crossclamp time for group III was 10 minutes. All anastomoses were patent at the time of 1-week reevaluation. Gross and microscopic examination demonstrated establishment of an intimal lining; organized nonocclusive thrombus was present in only one anastomosis. We conclude that atrium-to-atrium anastomosis is not necessary for a successful single lung transplantation, and that transplantation of a single lobe is feasible. The best alternative is implantation of the pulmonary vein into the left atrium, which will easily allow use of the heart and both lungs from a single donor to different recipients. We have used this anastomosis in one patient without difficulty.

Arterial aneurysm model using laser energy.

Continuous wavelength laser energy can be used to perform arterial anastomoses, but all experimental series report an incidence of anastomotic aneurysm formation. To elucidate the mechanism of aneurysm production, controlled injuries of the arterial wall were created with a pulsed CO2 laser beam (40-50 mW). One carotid and one femoral artery of 10 New Zealand rabbits were injured with laser and the contralateral vessel was exposed surgically as a sham operation. At reoperation 8 to 11 weeks later, all 40 arteries were patent. None of the carotid shams, one carotid laser, two femoral shams, and eight femoral laser vessels (80%) were aneurysmal. Histologic examination revealed extensive medial necrosis with fragmentation of the internal elastic lamina in the area of these aneurysms. Femoral vessels were significantly smaller than carotids (P less than 0.001) and the high incidence of aneurysm formation in the former may be due to the relatively greater area of injury. This new model of aneurysm formation after laser injury suggests a need for further study prior to clinical application of this technology, especially in vessels smaller than 2 mm in diameter.