Serum cell-free DNA levels are a useful marker for extramammary Paget disease.

BACKGROUND: Although carcinoembryonic antigen (CEA) and cytokeratin 19 fragment (CYFRA) are useful markers for extramammary Paget disease (EMPD), serum CEA and CYFRA levels are not elevated in most patients with EMPD without metastasis. Cell-free (cf)DNA has attracted attention as an indicator of clinical conditions in several cancers. OBJECTIVES: To identify further useful biomarkers for the detection of EMPD, including early lesions, and to study the clinical implications of cfDNA in EMPD. METHODS: cfDNA were isolated from serum of patients with EMPD with and without metastasis, and from healthy volunteers. Serum extracts were amplified using polymerase chain reaction. RESULTS: Serum cfDNA levels were significantly elevated in patients with EMPD with or without metastasis compared with those in healthy controls. Serum cfDNA was a better diagnostic marker for the presence of EMPD than serum CYFRA. Moreover, the postoperative serum cfDNA levels were significantly lower than those from the preoperative samples, and the change in serum cfDNA levels reflected the clinical courses of patients with EMPD treated with chemotherapy. CONCLUSIONS: Taking the evidence together, serum cfDNA levels may be a useful marker for diagnosis and disease progression in EMPD. What's already known about this topic? Serum levels of carcinoembryonic antigen (CEA) and cytokeratin 19 fragment (CYFRA) are not elevated in most patients with extramammary Paget disease (EMPD) without metastasis. Cell-free (cf)DNA has attracted attention as an indicator of clinical conditions in several cancers. There are few reports of the clinical implications of cfDNA in dermatology. What does this study add? Serum cfDNA levels were significantly elevated in patients with EMPD with or without metastasis compared with those in healthy controls. Postoperative serum cfDNA levels were significantly lower than those from the preoperative samples. Changes in serum cfDNA levels reflected the clinical courses of patients with EMPD treated with chemotherapy. What is the translational message? Serum cfDNA levels in patients with EMPD are a useful marker for the detection of EMPD, including localized EMPD. Changes in serum cfDNA levels in an individual patient may reflect the clinical course of EMPD.

Emphysematous cystitis following brain contusion.

Emphysematous cystitis is a rare disorder most commonly seen in patients with urinary tract infection and diabetes mellitus. We present a nondiabetic case of a 46-year-old woman with this entity following brain contusion.

Testis sparing surgery for infantile synchronous bilateral teratoma of the testis.

We report a rare case of infantile synchronous bilateral testicular teratoma treated by testis sparing surgery. Preoperative ultrasonography, tumor marker status and intraoperative findings were suggestive of benign neoplasia. Under these circumstances we performed high orchiectomy for the left testis and testis sparing surgery for the right. Surgical treatment of teratoma in childhood is controversial, especially when it occurs bilaterally. In this report we discuss testis sparing surgery as the treatment for this disease.

Expansion of venous flaps: an experimental study in rats.

In this study, the effects of controlled tissue expansion on the survival of unipedicled venous flaps were investigated in rats. In three groups of Lewis rats, a 3 x 6 cm unipedicled Lateral thoracic venous flap was studied with group 1 as control group (n = 30) without any manipulation, group 2 (n = 30) where an expander was introduced under the flap area but was not expanded and group 3 (n = 30) in which a 40 ml expander was introduced under the flap area and was expanded over a period of 10 days. Observations included gross and histological examination, reverse microangiography, reverse flow resistance and percentage of survival of flaps. Group 3 (expanded) showed more angiogenesis, a better delineation on microangiography, lower reverse flow resistance and higher survival rates as compared to group 2 (non-expanded). In the control group the survival rate of flaps was the poorest and the flaps showed higher rates of reverse flow resistance. Microangiography was not successful in the control group. In a pilot project prior to this experiment microscopic examination of the Lateral thoracic vein in 10 Lewis rats had confirmed the presence of two or three valves in this vein.

Venous drainage through bone marrow after replantation: an experimental study.

Venous drainage is vital for successful replantation, but it is not always possible to reconstruct because of missing or damaged veins. We devised an experimental model to study venous drainage through bone marrow while the new subcutaneous venous system regenerated. Adult male Wistar rats were placed into three groups. Group A rats had their hindlimbs amputated at the lower leg, but the tibia and sural and saphenous artery connections were preserved. Group B rats were prepared the same as Group A, except that a step-cut osteotomy was performed in the tibia. The bone ends were then realigned and kept in place with stainless steel wire. Group C rats were prepared the same as Group B, except that the ends of the bone were not aligned. All unoperated limbs served as controls for evaluations of blood flow. Experimental limbs were evaluated for skin colour and viability, blood flow and dye injection. Skin colour was investigated daily. Blood flow was measured postoperatively during three phases: immediate (up to 1 h), early (from 1 h to 24 h), and late (from 1 day to 7 days after operation). Survival of limbs varied in Groups A and B, while all limbs in Group C necrosed by day 7. Blood flow was returning to near control (normal) levels by day 7 in Group A and B limbs. India ink was observed in the medullary cavity at day 7. After replantation, bone marrow plays a critical role in venous drainage until the subcutaneous venous drainage system regenerates.

The effects of tissue expansion on the hemodynamic and survival characteristics of reverse-flow island flaps: an experimental study in rabbits.

The effects of tissue expansion on the hemodynamic and survival characteristics of reverse-flow island skin flaps were investigated in New Zealand White rabbits. The animals were divided into 3 groups of 15: group I (control) had no surgery prior to flap elevation, group II (nonexpansion) had a noninflated expander, and group III (expansion) had an inflated expander of 80 ml. After 3 weeks of expansion, a reverse-flow island flap based on the distal saphenous pedicle was elevated. A series of hemodynamic studies was performed to test reverse venous flow--in particular, valve competence. Besides observing the reverse flow under an operating microscope, the changes in the intravenous pressure were measured at 0, 5, 15, 30, 60, and 120 minutes after flap elevation. Moreover, reverse-flow resistance (RFR) was measured in each group to test the competence of venous valves. At each time interval, the values of intravenous pressure were significantly lower (p < 0.01) in group III than in the groups I and II. However there was no statistically significant difference between group I and group II. The RFR was measured as 126.7 +/- 33.52 mmHg in group I, 59.3 +/- 29.86 mmHg in group II (p < 0.01), and 25.1 +/- 7.68 mmHg in group III (p < 0.01). Ten days after flap elevation the mean survival of group III (100%) was statistically higher than that of group I (57.4 +/- 18.3%; p < 0.01) and group II (81.6 +/- 12.8%; p < 0.05). These findings simply suggest that controlled tissue expansion improves retrograde venous drainage and increases the survival of reverse-flow island flaps in rabbits. Abnormal dilatation of the venous tree and incompetence of the venous valves seem to be the main factors in explaining the decrease in the values of RFR and intravenous pressure in the expanded flaps. The potential mechanisms to explain the effects of tissue expansion, and the clinical implications are discussed.

Three approaches for laparoscopic unroofing of simple and complicated renal cysts.

We report 2 cases of simple renal cysts which were marsupialized with 2 laparoscopic approaches involving either transperitoneal, with reflection of the colon medially or dissection through the mesocolon, and a case of a multilocular renal cyst which was treated by the retroperitoneal approach. Although laparoscopic unroofing of a renal cyst is a safe and effective alternative to open surgical techniques, the transperitoneal approach should only be used for simple renal cysts. The retroperitoneal approach for complicated renal cysts may be indicated if preoperative examinations exclude the possibility of malignancy.

Low levels of mRNA for dopamine D4 receptor in human cerebral cortex and striatum.

Significant densities of mRNA for the dopamine D4 receptor in cerebral cortex, particularly in frontal lobe, have been reported in rats and monkeys, supporting the D4 hypothesis in the pathology of schizophrenia. Using northern blot analysis and the competitive reverse transcription-polymerase chain reaction (RT-PCR) method, we determined the relative levels of D4 mRNA in human brain regions to clarify whether the cortical level is also higher in humans. Northern blot analysis revealed an unexpected profile of D4 mRNA in the brain. The detected mRNA size, 1.5 kb, was quite different from the 5.3 kb reported in human neuroblastoma SK-N-MC cells. Higher levels of D4 mRNA were detected not only in the mesolimbic system but also in the corpus callosum, spinal cord, medulla, and subthalamic nucleus. It was surprising that in the cerebral cortex regions as well as the striatum, D4 mRNA was hardly detected. The competitive RT-PCR revealed these relative densities to be at least three orders of magnitude lower than that of the striatal D2 receptor. Our results demonstrate a remarkable difference in cortical D4 mRNA density in humans compared with that in rats and monkeys. Furthermore, the mRNA distribution suggests that the higher density of D4-like binding sites reported recently in normal human striatum is not due to the D4 receptor.

A true osteomyocutaneous free-flap model in rats: the saphenous artery osteomyocutaneous flap.

A new composite free-flap model, namely, the saphenous artery osteomyocutaneous flap, is described in the rat. This is a true osteomyocutaneous flap composed of a skin island from the medial aspect of the lower leg, the gracilis and semitendinosus muscles, and a bone segment from the tibia based on the saphenous vascular pedicle. After anatomic studies in 10 rats, 20 flaps were transplanted microsurgically to the abdominal region in the recipient rats with a 90 percent success rate. In selected animals, arteriography was carried out. The results from gross observation of the flap survival and histologic and fluorochrome bone-labeling studies revealed a complete survival for each component of the flap in all animals with a patent vascular pedicle at 2 weeks after transfer. The model seems to be suitable for use in metabolic, vascular, and immunologic experimental studies on composite free flaps.

The rat saphenous flap: a fasciocutaneous free flap model without panniculus carnosus.

The rat saphenous flap is described as a new experimental model for free flap studies. This is a fasciocutaneous free flap based on the saphenofemoral vascular pedicle. The flap may include the entire medial aspect of the lower leg between the knee and ankle. Thirty flaps were harvested from 15 inbred rats. Each flap was transferred to the anterior neck of a recipient rat of the same inbred strain so that 15 flaps were vascularized free flaps using the standard end-to-end microvascular technique and the other 15 flaps were nonvascularized free grafts. All but two (technical failure) of the vascularized flaps showed complete survival, whereas all nonvascularized flaps completely necrosed 2 weeks after transfer. It was concluded that the rat saphenous flap has several advantages such as a long and consistent vascular pedicle, ease of harvest, and an all-or-none survival pattern. Furthermore, as a unique feature of this flap, histological analysis revealed that the rat saphenous flap is composed of the skin and underlying fascia without panniculus carnosus. We therefore suggest that the rat saphenous flap is the first true fasciocutaneous free flap model in the rat. In this paper, in addition to illustrating the anatomy of the saphenous vessels and describing a new fasciocutaneous free flap model based on these vessels, we have documented some anatomical details of the rat leg that have never been described in the literature related to the rat anatomy.

Reversal by a dihydropyridine derivative of non-P-glycoprotein-mediated multidrug resistance in etoposide-resistant human prostatic cancer cell line.

PURPOSE: We have isolated etoposide-resistant prostatic cancer cell lines, P/VP10 and P/VP20, to investigate the multidrug resistance (MDR) mechanism and to find MDR reversal agents. MATERIALS AND METHODS: We examined expression of MDR-related genes and screened reversal agents of MDR in P/VP20 cells. RESULTS: These cells demonstrated a non-P-glycoprotein (P-gp)-mediated MDR phenotype with overexpression of MDR-associated protein (MRP) mRNA due to MRP DNA amplification. A 1,4-dihydropyridine derivative, bis(4-pyridylmethyl)4-[2-(3-methyl-5,6- dihydro-1,4-dithiinyl)]-2,6-dimethyl-1,4-dihydropyridine-3,5-dicar boxylate (NIK250), was found to overcome MDR in P/VP20 cells. CONCLUSIONS: NIK250 might be useful in reversing MDR, which often develops during chemotherapy of advanced or hormone-resistant prostatic cancer.

Is bed isolation necessary during flap prefabrication? An experimental study in rats.

This study was designed to investigate whether isolation of the bed is necessary during flap prefabrication. A fasciovascular carrier was used to prefabricate a 3 x 8-cm epigastric flap in rats. The animals were divided into three groups of 42 each. During flap prefabrication, in group 1 (control), no bed isolation was done, whereas in groups 2 and 3, a thin silicone sheet was used to isolate the underlying bed from the flap and fasciovascular carrier partially or completely, respectively. In each group, 14 prefabricated flaps were raised at 3, 7, and 14 days after prefabrication. Flap survival was assessed at day 7 after flap elevation. In comparing to the survival rates of the prefabricated flaps for each time interval, there was no significant statistical difference between the control and experimental groups (p > 0.05). With further studies, including india ink injection study and microangiographical and histological examinations of the prefabricated flaps, it was concluded that, during flap prefabrication, isolation of the bed with a silicone sheet does not improve the survival of prefabricated skin flaps. Moreover, it may cause some complications, such as extensive seroma, infection, and exposure of the sheet, which further interfere with the evaluation of experimental results.

Cutaneous vascular anatomy of the thoracic region of the dorsum and its role in flap design in the rat.

The cutaneous vascular anatomy of the proximal one-half of the dorsum (the thoracic region) and its role in flap design was studied in the rat. The investigation included anatomical dissection, ink injection into the axial artery, and flap harvesting in live animals. Anatomical dissection and india ink injection of the thoracic region revealed that the skin derives its principal blood supply from the lateral thoracic artery. The cutaneous vascular territory of the lateral thoracic artery was defined as follows: the medial border, along the midline of the back from the level of the inferior angle of the scapula to the lower border of the last rib; the lateral border, anterior axillary line corresponding with the length of the medial border; the cephalic border, a line joining the medial and lateral border at the level of the inferior angle of the scapula; and the caudal border, a line corresponding to the lower border of the last rib. Bilateral vascular pedicle island cutaneous flaps were harvested in living rats based on the vascular territory demarcated by India ink injection. All flaps survived; hence, this flap is reliable, with consistent vascularity, and is easy to harvest, and therefore, can be used as a vascular pedicle experimental model to study flap hemodynamics.

A new experimental model: the vascular pedicle cutaneous flap over the dorsal aspect (flank and hip) of the rat.

Cutaneous vascular anatomy of the flank and the hip was studied in the rat. Investigation was done by anatomical dissection, ink injection to the axial artery and flap harvesting in living rats. Anatomical dissection and India ink injection revealed that the rat's flank and hip skin derives its blood supply from the iliac branch of the iliolumbar artery, which originates from the lateral aspect of the abdominal aorta. The vascular territory of the iliac branch of the iliolumbar artery was defined as follows: the medial border--along the midline of the back from the 12th rib to the proximal part of the tail; the lateral border-anterior axillary line corresponding with the length of the medial border; the cephalic border--a line joining the medial and lateral border at the level of the 12th rib; and the caudal border--the posterior lateral line of the hip. On the basis of the vascular territory demarcated by India ink injection, bilateral vascular pedicle island cutaneous flaps were harvested in living rats. Results showed total survival of all flaps. This flap is easily made, and it could be used as a reliable model for flap research.

Nutritional status and daily physical activity of handicapped students in Tokyo metropolitan schools for deaf, blind, mentally retarded, and physically handicapped individuals.

Heights, weights, and skinfold thicknesses of 2222 handicapped students aged 3-22 y were measured in the 1984 nutritional survey for handicapped students in Tokyo metropolitan schools for deaf, blind, mentally retarded, and physically handicapped individuals. Although delayed growth was most obvious in physically handicapped students, obesity was already prevalent in many different types of handicapped students, especially those who were mentally handicapped. To estimate daily physical activity, 473 males and 329 females wore a pedometer for 24 h. There were considerable differences in the mean pedometer scores among the four groups of students: deaf greater than blind = mentally retarded greater than physically handicapped. In the female students who could walk normally, pedometer scores were negatively related with both body mass index and percent body fat. The nutritional status in the handicapped students is discussed in relation to daily physical activity.

Confirmation of the enantiomers of 1-methyl-1,2,3,4-tetrahydroisoquinoline in the mouse brain and foods applying gas chromatography/mass spectrometry with negative ion chemical ionization.

With the aid of a new chiral derivatizing reagent and a sensitive and specific assay using capillary gas chromatography/negative ion chemical ionization mass spectrometry, the proportions of R and S enantiomers of 1-methyl-1,2,3,4-tetrahydroisoquinoline (1MeTIQ) in mammalian tissues and foods were studied. R- and S-1MeTIQ enantiomers derivatized with a chiral derivatizing reagent, perfluoro-2-propoxypropionylchloride, were clearly separated. The ratio of enantiomers was R/S = 0.24, 0.55 and 0.60 in wine, cocoa and mouse brain. S-1MeTIQ predominated in all samples. This result suggested that 1MeTIQ could be formed at least partially through an enzymatic mechanism.

Activation of branched-chain alpha-keto acid dehydrogenase complex by exercise: effect of high-fat diet intake.

The effect of exercise on the activity of branched-chain alpha-keto acid dehydrogenase complex in liver and muscle was studied in rats fed a high-fat (FAT) or a high-carbohydrate (CHO) diet. Both diet groups of rats were offered isoenergetic diets by a meal-feeding method and were trained by treadmill running. On the final day of the experiment, half of the rats in each diet group were exercised by 2 h of running just before they were killed. The activity state of the enzyme complex was elevated maximally by exercise in liver of rats fed the FAT diet but not in liver of rats fed the CHO diet, suggesting that catabolism of branched-chain amino acids in rat liver during exercise was enhanced by the FAT diet. The activity state of the enzyme complex in muscle was enhanced by exercise in both groups of rats, but a significant difference was not observed between the groups. The concentration of branched-chain amino acids was elevated in liver and muscle by exercise in both groups of rats, but the elevated levels in liver were lower in rats fed the FAT diet than in those fed the CHO diet. Serum branched-chain amino acid concentrations were significantly lower in rested rats fed the FAT diet than in those fed the CHO diet, and the leucine and isoleucine concentrations in the former were elevated by exercise, but the serum concentrations in the latter were not significantly affected by exercise. ATP and ADP concentrations in muscle were not significantly affected by either diet or exercise.(ABSTRACT TRUNCATED AT 250 WORDS)