Standing on the shoulder of giants: Tubed pedicle radial forearm flap reconstruction for cutis aplasia.

INTRODUCTION: Aplasia cutis congenita presents a reconstructive challenge. We report the use of a tube pedicle radial forearm flap for scalp resurfacing in a patient who previously had two failed free flaps. CASE REPORT: A young male patient with cutis aplasia presented with a large full thickness defect of his occiput. He had multiple episodes of recurrent infections and a history of numerous surgeries as a child including tissue expansions. As a young adult he had two failed free flap reconstructions. A CT angiogram failed to demonstrate any adequate bloody supply to the scalp that would be suitable for another free tissue transfer. We successfully performed a tube pedicled flap from his forearm achieving good functional and aesthetic results. CONCLUSION: Originally descried by Filatov, the tube pedicle flaps remains a useful salvage option in the armamentarium of modern-day reconstruction.

Fault modeling and functional test methods for digital microfluidic biochips.

Dependability is an important attribute for microfluidic biochips that are used for safety-critical applications, such as point-of-care health assessment, air-quality monitoring, and food-safety testing. Therefore, these devices must be adequately tested after manufacture and during bioassay operations. Known techniques for biochip testing are all function oblivious (i.e., while they can detect and locate defect sites on a microfluidic array, they cannot be used to ensure correct operation of functional units). In this paper, we introduce the concept of functional testing of microfluidic biochips. We address fundamental biochip operations, such as droplet dispensing, droplet transportation, mixing, splitting, and capacitive sensing. Long electrode actuation times are avoided to ensure that there is no electrode degradation during testing. The functional testing of pin-constrained biochips is also studied. We evaluate the proposed test methods using simulations as well as experiments for a fabricated biochip.

Defect-aware high-level synthesis and module placement for microfluidic biochips.

Recent advances in microfluidics technology have led to the emergence of miniaturized biochip devices, also referred to as lab-on-a-chip, for biochemical analysis. A promising category of microfluidic biochips relies on the principle of electrowetting-on-dielectric, whereby discrete droplets of nanoliter volumes can be manipulated using an array of electrodes. As chemists adapt more bioassays for concurrent execution on such ldquodigitalrdquo droplet-based microfluidic platforms, system integration, design complexity, and the need for defect tolerance are expected to increase rapidly. Automated design tools for defect-tolerant and multifunctional biochips are important for the emerging marketplace, especially for low-cost, portable, and disposable devices for clinical diagnostics. We present a unified synthesis method that combines defect-tolerant architectural synthesis with defect-aware physical design. The proposed approach allows architectural-level design choices and defect-tolerant physical design decisions to be made simultaneously. We use a large-scale protein assay and the polymerase chain reaction procedure as case studies to evaluate the proposed synthesis method. We also carry out simulations based on defect injection to evaluate the robustness of the synthesized biochip designs.

Parallel scan-like test and multiple-defect diagnosis for digital microfluidic biochips.

Dependability is an important attribute for microfluidic biochips that are used for safety-critical applications such as point-of-care health assessment, air-quality monitoring, and food-safety testing. Therefore, these devices must be adequately tested after manufacture and during bioassay operations. We propose a parallel scan-like testing methodology for digital microfluidic devices. A diagnosis method based on test outcomes is also proposed. The diagnosis technique is enhanced such that multiple defect sites can be efficiently located using parallel scan-like testing. Defect diagnosis can be used to reconfigure a digital microfluidic biochip such that faults can be avoided, thereby enhancing chip yield and defect tolerance. We evaluate the proposed method using complexity analysis as well as applying it to a fabricated biochip.

Induction of uterine prostaglandin H synthase 2 by estradiol following fetal adrenalectomy.

In sheep, fetal cortisol stimulates the conversion of progesterone to estradiol in late gestation initiating labor. It is unclear whether an intact fetal hypothalamic-pituitary-adrenal (HPA) axis is required to induce the estradiol-triggered subsequent endocrine changes including enhanced intrauterine prostaglandin (PG) synthesis associated with the onset of labor. We have shown that maternal estradiol administration stimulates PG H synthase (PGHS)-2 expressions in pregnant ovine intra-uterine tissues. The current study was undertaken to determine whether the fetal adrenal mediates estradiol's stimulation of the intrauterine PGHS-2 in pregnant sheep. Placenta, myometrium, and endometrium were collected from two groups of ewes at 123-127 d of gestational age (dGA) after fetal adrenalectomy and vehicle treatment (ADX; n = 5); or fetal ADX and maternal estradiol administration (5 mg twice a day for 2 d, ADX+E2, n = 5). PGHS-2 mRNA and protein were analyzed by Northern and Western Blot analyses in both groups and presented as the ratios to beta actin mRNA for Northern and G protein beta subunit for Western blot analysis. Fetal plasma cortisol was measured by radioimmunoassay. Data were analyzed by Student's t test. Fetal plasma cortisol levels were low in ADX and ADX+ E2 groups (<6 ng/mL). The cervix of all ADX+E2 treated ewes was dilated at necropsy. Three out of five ADX+ E2-treated ewes delivered within 48 h. The cervix was closed in all fetal ADX ewes at necropsy. PGHS-2 mRNA and protein increased (p < 0.05) in myometrium and endometrium, but not placenta in ADX+E2-treated ewes compared with ADX group. These data provide the first in vivo evidence for estradiol upregulation of intrauterine PGHS-2 in late gestation in the absence of an intact fetal HPA axis. Thus, the fetal adrenal is not required to mediate estradiol's stimulation of uterine PGHS-2 expression associated with the onset of labor.

Distally based dorsal metacarpal flaps: a review of a series of patients treated in a 6-month period.

The management of dorsal digital and hand injuries can pose a difficult problem to the trainee surgeon. Often repair involves using tissue from neighbouring digits. Our experience using local perforator flaps based on the direct cutaneous branch of the dorsal metacarpal arteries shows that these can be dissected and used successfully in the management of complex injuries of the dorsum of the hand and proximal phalanges. The flap has been used in a series of seven patients treated over a 6-month period by plastic surgery trainees with 100% success. This one-stage procedure resulted in good functional and aesthetic outcome and left an acceptable donor scar.

Characterization of two labor-induced genes, DSCR1 and TCTE1L, in the pregnant ovine myometrium.

In the present study we characterized two labor-induced genes, DSCR1 (Down syndrome candidate region 1) and TCTE1L (murine t-complex like), which were identified by suppression subtractive hybridization in the pregnant ovine myometrium. DSCR1 and TCTE1L cDNA sequences were retrieved from a custom-made labor-myometrial cDNA library by hybridization screening. The characterized cDNA sequences include 5'-untranslated region (UTR), coding region and 3'-UTR, which are 12 bp, 351 bp and 1716 bp for TCTE1L, and 64 bp, 594 bp and 1539 bp for DSCR1 respectively. The two cDNA sequences encode proteins of 116 and 197 amino acids for TCTE1L and DSCR1 respectively. Northern analysis further confirmed the significant increases of myometrial DSCR1 and TCTE1L mRNA associated with spontaneous term labor (n=6) compared with gestation-matched controls not in labor (n=6). The abundance of DSCR1 and TCTE1L mRNA was attenuated when myometrial contraction was inhibited by Nimesulide (n=6), a specific prostaglandin H synthase 2 inhibitor. Fetal occupancy greatly upregulated DSCR1 and TCTE1L mRNA in the gravid horn during betamethasone-induced premature labor (n=6) compared with the non-gravid horn not in labor (n=3). Estradiol upregulated TCTE1L mRNA, but had no effect on DSCR1 mRNA expression in the non-pregnant sheep myometrium. Progesterone alone had no effect on both DSCR1 and TCTE1L mRNA expression, however progesterone antagonized estradiol's stimulating effect on myometrial TCTE1L mRNA expression in ovariectomized non-pregnant sheep. Upregulation of DSCR1 and TCTE1L in both betamethasone-induced premature labor and spontaneous term labor and inhibition of their expression by Nimesulide suggest a functional role of these two genes in myometrial activation associated with onset of labor. Mechanical stretch, labor and steroids differentially regulated DSCR1 and TCTE1L mRNA in the pregnant and non-pregnant sheep myometrium.

Chaos in the brain: a short review alluding to epilepsy, depression, exercise and lateralization.

Electroencephalograms (EEGs) reflect the electrical activity of the brain. Even when they are analyzed from healthy individuals, they manifest chaos in the nervous system. EEGs are likely to be produced by a nonlinear system, since a nonlinear system with at least 3 degrees of freedom (or state variables) may exhibit chaotic behavior. Furthermore, such systems can have multiple stable states governed by "chaotic" ("strange") attractors. A key feature of chaotic systems is the presence of an infinite number of unstable periodic fixed points, which are found in spontaneously active neuronal networks (e.g., epilepsy). The brain has chemicals called neurotransmitters that convey the information through the 10(16) synapses residing there. However, each of these neurotransmitters acts through various receptors and their numerous subtypes, thereby exhibiting complex interactions. Albeit in epilepsy the role of chaos and EEG findings are well proven, in another condition, i.e., depression, the role of chaos is slowly gaining ground. The multifarious roles of exercise, neurotransmitters and (cerebral) hemispheric lateralization, in the case of depression, are also being established. The common point of reference could be nonlinear dynamics. The purpose of this review is to study those nonlinear/chaotic interactions and point towards new theoretical models incorporating the oscillation caused by the same neurotransmitter acting on its different receptor subtypes. This may lead to a better understanding of brain neurodynamics in health and disease.

Keratinocyte-driven contraction of reconstructed human skin.

We have previously reported that reconstructed human skin, using deepidermized acellular sterilized dermis and allogeneic keratinocytes and fibroblasts, significantly contracts in vitro. Contracture of split skin grafts in burns injuries remains a serious problem and this in vitro model provides an opportunity to study keratinocyte/mesenchymal cell interactions and cell interactions with extracted normal human dermis. The aim of this study was to investigate the nature of this in vitro contraction and explore several approaches to prevent or reduce contraction. Three different methodologies for sterilization of the dermal matrix were examined: glycerol, ethylene oxide and a combination of glycerol and ethylene oxide. While the nature of the sterilization technique influenced the extent of contraction and thinner dermal matrices contracted proportionately more than thicker matrices, in all cases contraction was driven by the keratinocytes with relatively little influence from the fibroblasts. The contraction of the underlying dermis did not represent any change in tissue mass but rather a reorganization of the dermis which was rapidly reversed (within minutes) when the epidermal layer was removed. Pharmacological approaches to block contraction showed forskolin and mannose-6-phosphate to be ineffective and ascorbic acid-2-phosphate to exacerbate contraction. However, Galardin, a matrix metalloproteinase inhibitor and keratinocyte conditioned media, both inhibited contraction.

The reversible antifertility effect of Piper betle Linn. on Swiss albino male mice.

To study the antifertility effect of an extract (alcoholic) of the leaf-stalk of Piper betle Linn., one set of experiments with two different doses in Swiss male albino mice were evaluated. Initially, 500 mg of the leaf-stalk extractive for 30 days and then 1000 mg for next 30 days/animal/day/kg body weight were administered orally. The extract reduced fertility to 0% within 60 days. Suppression of cauda epididymal sperm count and motility (p <0.05) was observed. Biochemical parameters did not show any marked alterations in testosterone content in serum nor 17beta-hydroxysteroid dehydrogenase (17beta-HSD) activity in testes although fructose content in seminal vesicles was reduced as are the weights of reproductive organs. The cholesterol content in testes increased, although not appreciably. After cessation of drug (plant extract) treatment, the altered parameters recovered. Results suggest that the contraceptive effect of the extract of leaf-stalk of Piper betle Linn. is mainly on the maturation process of spermatozoa in epididymides without influencing hystemic hormonal profiles. Withdrawal of the extract restored all altered parameters including organ weights and fertility after 60 days.

Development of autologous human dermal-epidermal composites based on sterilized human allodermis for clinical use.

The aim of this study was to identify a sterilization technique for the preparation of human allodermis which could be used as a dermal component in wound healing and as the dermal base for production of dermal-epidermal composites for one-stage grafting in patients. We report that it is possible to produce dermal-epidermal composites which perform well in vitro and in vivo using a standard ethylene oxide sterilization methodology. Prevention of ethylene oxide-induced damage to the dermis was achieved using gentle dehydration of the skin prior to ethylene oxide sterilization. The issue of whether viable fibroblasts are required for composite production was examined in comparative studies using glycerol vs. ethylene oxide sterilized dermis. Where good collagen IV retention was achieved following preparation of acellular de-epidermized dermis there was no advantage to having fibroblasts present in vitro or in vivo; however, where collagen IV retention was poor or where keratinocytes were initially expanded in culture then there was a significant advantage to introducing fibroblasts to the composites during their preparative 10-day period in vitro. The requirement for fibroblasts became less evident when composites were grafted on to nude mice. In conclusion, we report a protocol for the successful sterilization of human allodermis to achieve an acellular dermis with good retention of collagen IV. This acellular dermis would be appropriate for clinical use as a dermal replacement material. It can also be used for the production of dermal-epidermal composites using autologous keratinocytes (with or without fibroblasts).

Immobilisation stress induced analgesia in diabetic rats.

Stress is known to produce analgesia. The pain threshold is altered in diabetes. We studied the effect of 1 hr of immobilisation stress on pain threshold in male Wistar rats. The same effect was tested in streptozotocin induced diabetic rats. The pain threshold of tail flick, vocalisation and vocalisation after discharge increased in the control group after the stress procedure. Significant analgesia was also obtained in diabetic rats, for flick and after discharge pain threshold. However the vocalisation threshold was not altered, probably due to the antagonistic action of glucose on opiate receptor at the level of brain stem.

Nociception, antinociceptive potency of morphine in streptozotocin induced diabetic rats.

There are controversial reports on the effect of diabetes on the pain threshold. We used male Wistar rats to see the effect of streptozotocin induced diabetes on the tail flick, vocalisation and vocalisation after discharge responses. These represent the spinal, lower brain stem and hypothalamic responses respectively. The effect of morphine in these parameters was studied for both the control and diabetic group. In diabetic rats, the pain threshold was increased. However, this increase was not significant. Morphine produced significant analgesia after thirty minutes for tail flick and vocalisation responses and after fifteen minutes for after discharge in the control group. The antinociceptive effect of morphine was delayed and reduced for all three pain threshold confirming the antagonistic action of glucose on opiate receptors.

Ferguson-Smith syndrome: the importance of long term follow-up.

A case is presented which highlights the importance of long term follow-up and early intervention in this unusual dermatosis. A 39-year-old man presented with an extensive tumour of his left ear and scalp. Surgery was not feasible. The progressive nature of the lesion eventually killed the patient.

Hypotensive effect of intracerebroventricular injection of norepinephrine and its modulation by alpha and beta adrenergic blockers in conscious rabbits.

The present study was designed to investigate the role of central adrenoceptors in the hypotensive effect of intracerebroventricular (ICV) injection of norepinephrine (NE) in conscious rabbits. Experiments were carried out on 19 adult rabbits (oryctolagus cuniculus) of either sex. A dose-dependent hypotensive response to ICV injection of NE was observed with no significant change in heart rate. The hypotensive response of NE was blocked 74.2 +/- 0.7% by yohimbine (alpha-2 adrenergic blocker), and 25.0 +/- 0.5% by metoprolol (beta-1 adrenergic blocker). NE response was not affected either by prazosin or butoxamine (alpha-1 and beta-2 adrenergic blockers respectively). The results suggest that the dose-dependent hypotensive response of ICV administered NE is mediated through alpha-2 and beta-1 central adrenoceptors.

Characterization of guanosine diphospho-D-mannose dehydrogenase from Pseudomonas aeruginosa. Structural analysis by limited proteolysis.

Alginate is believed to be a major virulence factor in the pathogenicity of Pseudomonas aeruginosa in the lungs of patients suffering from cystic fibrosis. Guanosine diphospho-D-mannose dehydrogenase (GDPmannose dehydrogenase, EC 1.1.1.132) is a key enzyme in the alginate biosynthetic pathway which catalyzes the oxidation of guanosine diphospho-D-mannose (GDP-D-mannose) to GDP-D-mannuronic acid. In this paper, we report the structural analysis of GMD by limited proteolysis using three different proteases, trypsin, submaxillary Arg-C protease, and chymotrypsin. Treatment of GMD with these proteases indicated that the amino-terminal part of this enzyme may fold into a structural domain with an apparent molecular mass of 25-26 kDa. Multiple proteolytic cleavage sites existed at the carboxyl-terminal end of this domain, indicating that this segment may represent an exposed region of the protein. Initial proteolysis also generated a carboxyl-terminal fragment with an apparent molecular mass of 16-17 kDa which was further digested into smaller fragments by trypsin and chymotrypsin. The proteolytic cleavage sites were localized by partial amino-terminal sequencing of the peptide fragments. Arg-295 was identified as the initial cleavage site for trypsin and Tyr-278 for chymotrypsin. Catalytic activity of GMD was totally abolished by the initial cleavage. However, binding of the substrate, GDP-D-mannose, increased stability toward proteolysis and inhibited the loss of enzyme activity. GMP and GDP (guanosine 5'-mono- and diphosphates) also blocked the initial cleavage, but NAD and mannose showed no effect. These results suggest that binding of the guanosine moiety at the catalytic site of GMD may induce a conformational change that reduces the accessibility of the cleavage sites to proteases. Binding of [14C]GDP-D-mannose to the amino-terminal domain was not affected by the removal of the carboxyl-terminal 16-kDa fragment. Furthermore, photoaffinity labeling of GMD with [32P]arylazido-beta-alanine-NAD followed by proteolysis demonstrated that the radioactive NAD was covalently linked to the amino-terminal domain. These observations imply that the amino-terminal domain (25-26 kDa) contains both the substrate and cofactor binding sites. However, the carboxyl-terminal fragment (16-17 kDa) may possess amino acid residues essential for catalysis. Thus, proteolysis had little effect on substrate binding, but totally eliminated catalysis. These biochemical data are in complete agreement with amino acid sequence analysis for the existence of substrate and cofactor sites of GMD. A linear peptide map of GMD was constructed for future structure/functional studies.

Effect of cyproheptadine on glucose tolerance, serum insulin and structure of pancreatic islets in rats.

The effect of cyproheptadine (CPH) on glucose tolerance, serum immunoreactive insulin (IRI) and structure of pancreatic islets in albino rats has been studied. Hyperglycemia with glucose intolerance was observed after 10 days of administration of CPH (40 mg/kg, ip). There was insignificant change of fasting IRI after the treatment. Histological studies indicated degranulation and vacuolation of beta cells with enlargement of capillaries. Improvement in blood glucose, glucose tolerance and structure of islets with proliferation of small pancreatic ducts and cell cords were observed 10 days after the withdrawal of CPH.