Human Sgo1 downregulation leads to chromosomal instability in colorectal cancer.

BACKGROUND AND AIMS: Chromosomal instability (CIN) is recognised as a hallmark of cancer and is caused by a spindle assembly checkpoint disorder or chromosome mis-segregation during mitosis. Although the recent identification of human shugoshin (hSgo1), an important player in proper chromosome segregation, has suggested the involvement of hSgo1 in colorectal tumourigenesis, little is known about how it is involved. The aim of this study was to obtain information about the status of hSgo1 in human colorectal cancer. METHOD AND RESULTS: Among the 46 colorectal cancer cases, hSgo1 mRNA expression was decreased in the tumour tissue in comparison with the corresponding normal tissue (p = 0.032). Human Sgo1-downregulated tumours (tumour to normal mucosa ratio<0.5) had preferential location on the left side large bowel rather than on the right side (p = 0.012), and a higher variation of centromere numbers revealed by fluorescence in situ hybridisation (FISH). To assess the effects of hSgo1 downregulation, hSgo1 knockdown was performed by transfecting the diploid HCT116 cell line with a short hairpin RNA expression vector. hSgo1 knockdown cells proliferated slowly because of both G(2)/M arrest and apoptosis (p<0.001), and markers of CIN in the form of aneuploidy (p<0.001) and micronuclei (p<0.005) were later observed in hSgo1 knockdown cells. Increased centrosome amplification (p<0.05), the presence of binucleated cells and mitotic catastrophes were also noted in hSgo1 knockdown cells. CONCLUSIONS: These findings suggest that hSgo1-downregulated colorectal cancers have a clinicopathological character of CIN, and hSgo1 downregulation leads to CIN in colorectal cancer cells.

Size-dependent uptake of electrically neutral amphipathic polymeric nanoparticles by cell-sized liposomes and an insight into their internalization mechanism in living cells.

Size-dependent uptake behaviors of electrically neutral amphipathic polymeric nanoparticles in cell-sized liposomes and living cells were investigated. Kinetic analyses and the particle size distribution suggested a size-dependent penetration mechanism (size threshold: 3.1 nm). The definite size-dependent uptake provides a new insight into the interactions between nanomaterials and living cells.

Malonyl-CoA: acetyl-CoA cycling. A new micromethod for determination of acyl-CoAs with malonate decarboxylase.

A new micromethod for determination of acetyl-CoA and malonyl-CoA using malonate decarboxylase is described. This enzyme catalyzes decarboxylation of malonate in a cyclic manner and produces acetate in proportion to the amount of a given acyl-CoA, such as acetyl-CoA, malonyl-CoA or propionyl-CoA. The acetate generated is converted to acetylphosphate by acetate kinase (EC 2.7.2.1) added at the same time and is determined spectrophotometrically as acetohydroxamate. The sensitivity of this method is high enough to detect 10(-12) mol of acetyl-CoA or malonyl-CoA. The simplicity of the method allows more than 30 samples to be analyzed at the same time without any prior extraction step. Although this method does not distinguish between acetyl-CoA and malonyl-CoA, malonyl-CoA alone can be measured by elimination of acetyl-CoA with citrate synthetase (EC 4.1.3.7).

Different vulnerability among chromosomes to numerical instability in gastric carcinogenesis: stage-dependent analysis by FISH with the use of microwave irradiation.

Numerical chromosomal abnormalities are a well known characteristic of human cancer, but no "chromosome-wide investigation" encompassing almost all of the chromosomes has ever been reported. Furthermore, although the multistep process of carcinogenesis is widely accepted in human cancer, the stepwise numerical aberration of chromosomes has never been addressed. Touch preparations of 24 (male 20, female 4) surgically resected gastric cancer tissue samples in various stages in terms of depth of invasion were analyzed by fluorescence in situ hybridization using centromere-specific probes including 17 chromosomes, 1-4, 6-8, 10-12, 15-18, 20, X, and Y. Microwave irradiation was performed to increase the sensitivity and specificity of the signal. The depth of the tumor invasion in the gastric wall and histological subtypes were recorded by viewing the histology of the adjacent portion. Numerical chromosomal abnormalities of chromosomes 1 and 2 were found most frequently and from the early stage of gastric cancer. The abnormalities observed were limited to chromosomes 1, 2, 4, and 20 in tumors invading to the middle layer of the submucosa of the gastric wall, but these became more extensive, involving almost all of the chromosomes investigated when the tumor had invaded beyond the proper muscle of the gastric wall. Centromeric numbers of chromosomes 3 and 18 were exceptionally stable even after the tumor progressed to advanced stage. These profiles of the sequential process of numerical chromosomal abnormality were similar in both mucocellular and tubular-type gastric cancer, but the prevalence was significantly lower in the mucocellular type (39.0% versus 68.0%). On the basis of fluorescence in situ hybridization analysis of 17 different chromosome centromeres in gastric cancer in various stages, we conclude that the earliest events in gastric carcinogenesis in terms of chromosomal abnormality occur in chromosomes 1 and 2 and that chromosomal numerical aberrations expand in a stepwise manner with cancer progression.

Alteration of immunoreactivity by hydrated autoclaving, microwave treatment, and simple heating of paraffin-embedded tissue sections.

The effects of treatment in a hydrated autoclave (121 degrees C, 2 atm for 20 min), microwave oven (in water), and simple heating (60 degrees C overnight in distilled water or 90 degrees C for 10 min in ZnSO4) on the stainability of 56 antigens by commercially available antibodies in formalin-fixed paraffin-embedded tissue sections were evaluated. The detectability of nuclear antigens, glycoprotein, lymphocytic surface markers, and chromogranin A was significantly and reproducibly improved by these treatments, whereas the detectability of viral antigens and peptide hormones was attenuated or unchanged. This enhancement includes not only the distinctiveness of the positive staining, but also the number of positive cells, as revealed by comparing serial sections. Among these four heating procedures, microwave heating and autoclaving were more effective than the others on p53, c-erbB-2, and CA125, whereas simple heating was best for smooth-muscle actin (HHF35 and CGA7). Generally the effects of the heating procedures for these antigens were consistent among the cases, but the effects on GFAP varied with the case. The alterations we observed could significantly influence the interpretation of immunohistochemical staining of currently popular tumor markers such as p53 in terms of their prevalence (28% vs 64% in gastric cancer; 36% vs 82% in metastatic liver cancer) and other diagnostically important markers.

Expression of p53 and flow cytometric DNA analysis of isolated neoplastic glands of the stomach: an application of the gland isolation method.

The expression of p53 was studied immunohistochemically in combination with the DNA ploidy pattern by gland isolation in 97 alcohol-fixed gastric lesions. A polyclonal antibody, CM-1, was applied to the paraffin-embedded sections in this study. Overexpression of the p53 protein was found in 73.2% of 41 well or moderately differentiated gastric carcinomas and 52.2% of 23 cases with poor differentiation (P < 0.05). Immunoreactivity of p53 was also detected in isolated cancerous glands. No p53 immunoreactivity was detected in benign gastric lesions including adenomas, hyperplastic polyps and regions of intestinal metaplasia. In addition, flow cytometric DNA analysis was performed on isolated glandular epithelium adjacent to the portions used for immunostaining. DNA aneuploidy (DA) was detected in 85.7% of the well or moderately differentiated carcinomas and 42.9% of those with poor differentiation (P < 0.05). There was a positive correlation between DA, p53 positivity and the presence of regional lymph node metastasis, but not with other clinicopathological variables. In spite of the limited applicability of this method to poorly differentiated gastric cancer, we found that immunostaining and flow cytometry in combination with the gland isolation method facilitates analysis of gastric carcinogenesis.

Congenital onychodysplasia. Report of 11 cases.

We have seen 11 cases of congenital onychodysplasia over the past six years. Associated anomalies were present in six patients, most of whom had an anomaly of the hand. Roentgenograms of the affected fingers showed hypoplasia, narrowing at the distal third of the distal phalanx, and bifurcation of the distal phalanx. The etiology of this condition is unknown, but clinical study suggested that ischemia of the finger at a certain period of embryonic life might play an important part in its pathogenesis.

Gastric plasmacytoma coexistent with plasma cell granuloma in a patient with gastric cancer.

We report a rare case of multiple gastric plasmacytomatous lesions at an early stage found incidentally in a stomach resected because of gastric cancer. The three lesions had different macroscopic features, showing depressed, submucosal, and nodular tumor-forming types. The smallest, a depressed lesion, produced IgG of the lambda and kappa types, and had a plasma-cell granuloma-like appearence, whereas the largest, a submucosal tumor, was formed by the monoclonal proliferation of atypical plasma cells containing IgA-kappa type immunoglobulin, and the nodular lesion exhibited histological features intermediate between those of the other two lesions. From their microscopic features and the profile of immunoglobulin production, we believe that these lesions may represent different stages in the possible course of development of plasmacytoma from the early stage of plasma cell granuloma. All three lesions were located far from the gastric carcinoma and it was unclear whether they had any causal relationship with it.

Patterns of arterial distribution in the duplicated thumb.

The arterial blood supply of the duplicated thumb was studied in 42 hands using the angiography technique. The developing factor of the duplicated thumb also may hinder the developing process of the limb artery, especially in the early stage when the median artery develops. In 73.8 percent of duplicated thumbs, two digital arteries were present, and 96.8 percent of them had one digital artery in each member. Simple ablation of the supernumerary digit leaves the thumb with a single end-arterial blood supply. The developmental mechanism of the duplicated thumb may depend not on duplication of the limb bud, but on splitting of the limb mesenchyme. The group of type VII of Wassel's classification was different from the other types in terms of origin of the digital artery distributing blood to the duplicated thumb.

Usefulness of cord-blood harvesting for autologous transfusion in surgical newborns with antenatal diagnosis of congenital anomalies.

BACKGROUND/PURPOSE: The risks of homologous transfusion and the effectiveness of predeposit autologous transfusion have been described. The authors examined the clinical usefulness of cord-blood harvesting for autologous transfusion in newborns who had congenital anomalies antenatally diagnosed that would require surgical intervention at or near the time of delivery. METHODS: Of 112 cases of antenatal diagnosis of congenital anomalies, 50 mothers gave informed consent and enrolled in this study. Cord-blood was withdrawn immediately after clamping of the umbilical cord and was used for autologous transfusion in newborns within the first 3 days postpartum. RESULTS: A mean of 72 +/- 54 mL of cord-blood was harvested (27 +/- 18 mL/kg). While preserving cord-blood for 3 days at 4 degrees C, no signs of clot formation or hemolysis were observed. The harvested cord-blood included plasma-free Hb ranging from 1 to 68 (13 +/- 18) mg/dL and thrombin-antithrombin III complex ranging from 2 to 273 (18 +/- 50) ng/mL. Bacteriologic examination of the stored cord-blood showed negative cultures, except for samples from 3 newborns after vaginal delivery. A mean of 46 +/- 34 mL of cord-blood was used in 26 patients for autologous transfusion. No significant complications related to cord-blood transfusion were recognized clinically. CONCLUSIONS: Autologous cord-blood transfusion has the potential to be a useful alternative to homologous transfusion in newborns requiring surgery. Adequate collection and storage techniques for cord-blood must be developed. J Pediatr Surg 36:851-854.

Experience in tracheobronchial reconstruction with a costal cartilage graft for congenital tracheal stenosis.

Although successful surgical management of congenital tracheal stenosis has been reported, it is still controversial as to the best operative procedure. Eleven infants with congenital tracheal stenosis were evaluated to confirm the efficacy of tracheobronchial reconstruction with costal cartilage graft. Symptoms ranged from recurrent respiratory infection to severe respiratory failure. All infants had other congenital anomalies in addition to tracheal stenosis. Notably, five infants had pulmonary artery sling and four infants had patent ductus arteriosus. Definitive diagnosis was made by bronchoscopy, results of which showed complete tracheal rings in all patients with severely compromised tracheobronchial lumens. Five infants had elongated stenosis involving nearly the whole length of the trachea, and five infants had segmental stenosis involving nearly one half the length of the trachea. One infant had bilateral stenosis of the main bronchi. Early experience included two deaths from problems related to the repair. The involvement of the carina and the distal portion of the trachea was associated with increased complications and a higher mortality rate. Currently, our preferred technique facilitated by extracorporeal membrane oxygenation (ECMO) includes carinal reconstruction with a thin-wall intraluminal stent. Bronchoscopy is essential for accurate intraoperative incision of the trachea, post-operative airway management for several weeks, and removal of the intraluminal stent.

Nitric oxide inhalation therapy for an infant with persistent pulmonary hypertension caused by misalignment of pulmonary veins with alveolar capillary dysplasia.

Misalignment of pulmonary veins with alveolar capillary dysplasia (MPV) has been reported to be a rare cause of persistent pulmonary hypertension of the newborn (PPHN) and to be fatal despite extracorporeal membrane oxygenation (ECMO). A full-term female neonate with PPHN was brought to the hospital for ECMO therapy at 2 days of age. On the 14th day of life, she was extubated early after the second run of ECMO, and underwent nitric oxide (NO) inhalation therapy in the incubator. She died of catheter-related sepsis on the 61st day of life. After autopsy findings revealed MPV, the longest survival with this disease was documented. NO inhalation therapy in the incubator may provide time for lung transplantation.

Patent ductus venosus with a hypoplastic intrahepatic portal system presenting intrapulmonary shunt: a case treated with banding of the ductus venosus.

A case of patent ductus venosus (PDV) presenting intrapulmonary shunting is described. Although retrograde venography of ductus venosus showed few intrahepatic branches, banding of PDV resulted in increased intrahepatic portal branches and disappearance of symptoms 10 months after the operation. Banding of the ductus venosus may be effective in PDV even with hypoplastic intrahepatic portal system.

Pexis of the great vessels for patients with tracheobronchomalacia in infancy.

BACKGROUND/PURPOSE: Aortopexy has been established as a surgical treatment for tracheobronchomalacia in infancy. However, the effects in patients with bronchial involvement remain controversial. The authors performed aortopexy and pexis of the pulmonary artery trunk (PApexy) for patients with tracheobronchomalacia. METHODS: Between 1992 and 1997, 14 infants with tracheobronchomalacia were treated. Patients ranged in age from 4 months to 7 years (median, 3.5 years). Using patients' records, intraoperative bronchofiberscopic results and clinical outcomes were analyzed retrospectively. RESULTS: Concerning tracheomalacia, aortopexy eliminated airway collapse as shown by intraoperative fiberscopic results and clinical outcome in 4 patients. Regarding tracheobronchomalacia, aortopexy eliminated collapse of the trachea and the right main bronchus in 3 patients but did not improve collapse of the lower half of the left main bronchus in 3 patients, which was eliminated by PApexy. Aortopexy eliminated airway collapse of the left main bronchus in 1 of 3 patients with bronchomalacia. PApexy eliminated collapse of the lower half of the left main bronchus in 1 patient. CONCLUSIONS: These results indicate that intraoperative bronchofiberscopic results were closely correlated to the clinical outcome, and suggest that aortopexy is helpful for eliminating collapse of the right main bronchus and the proximal half of the left main bronchus and that PApexy eliminates collapse of the distal half of the left main bronchus.

Prenatal diagnosis of abdominal wall defects and their prognosis.

To determine whether associated anatomic features may be useful in predicting fetal prognosis, 43 fetuses who had ultrasonographic evaluation for abdominal wall defects during a 13-year period were reviewed. Thirty-one fetuses had omphalocele and were classified into three groups according to fetal ultrasonography results: ruptured omphalocele with exposed liver, giant omphalocele with exposed liver, and small omphalocele without liver herniation. Twelve fetuses had gastroschisis. Ten of the 12 fetuses with gastroschisis survived; one died in utero. Nine of the 12 with a small omphalocele survived. Ten of 12 fetuses with giant omphalocele survived, but six of the seven with ruptured omphalocele died of pulmonary hypoplasia and respiratory insufficiency. Ruptured omphalocele recognized in utero was accompanied by intrauterine growth retardation and liver herniation and frequently was associated with deformity of the spine, diaphragmatic defects, vesicointestinal fissure, and meningocele. These results indicated that, together with the deformity of the spine, rupture and absence of the covering membrane with an exposed liver (noted through fetal ultrasonography) may suggest a poor prognosis because of pulmonary hypoplasia.

Effects of sevoflurane anaesthesia on renal function.

The effects of sevoflurane on renal function were investigated in 34 patients anaesthetized with sevoflurane. Based on preoperative serum creatinine levels, patients were classified into three groups: Group 1 (n = 25) had normal renal function (serum creatinine < 1.0 mg/dl); Group 2 (n = 5) had slight renal dysfunction (1.0 < or = serum creatinine < 1.5 mg/dl); Group 3 (n = 4) had moderate renal dysfunction (serum creatinine > or = 1.5 mg/dl]. Serum creatinine, blood urea nitrogen and urine volume were measured preoperatively, and at Days 0, 1, 2, 7 and 28 after operation. Serum creatinine and blood urea nitrogen showed no significant postoperative differences (P < 0.05) in each group, whereas urine volume showed a significant increase until Day 2, with no further changes thereafter. Our results suggest that sevoflurane anaesthesia causes no significant renal damage in patients with normal and insufficient renal function under normal-duration anaesthesia within 3-4 h.

Reducing nurses'. Workload using a computerized nursing support system linked to the hospital information system.

A computerised nursing support system (CNSS) linked to the hospital information system (HIS) was developed and has been in use for one year, in order to reduce the workload of nurses. CNSS consists of (1) a hand held computer for each nurse (2) desk-top computers in the nurses' station and doctors' rooms (3) a data server (4) an interface with the main hospital information system. Nurses enter vital signs, food intake and other information about the patients into the hand held computer at the bed-side. The information is then sent automatically to the CNSS data server, which also receives patients' details (prescribed medicines etc.) from the HIS. Nurses and doctors can see all the information on the desk-top and hand held computers. This system was introduced in May 1995 into a university hospital ward with 40 beds. A questionnaire was completed by 23 nurses before and after the introduction of CNSS. The mean time required to post vital data was significantly reduced from 121 seconds to 54 seconds (p < 0.01). After three months 30% of nurses felt CNSS had reduced their workload, while 30% felt it had complicated their work; after five months 70% noted a reduction and 0% reported that CNSS had made their work more complex. The study therefore concludes that the interface between a computerised nursing support system and the hospital information system reduced the workload of nurses.

High division of the median nerve: unusual anatomical variation.

An unusually high division of the median nerve in the forearm is reported. Recognition of this anatomical variation can help plastic surgeons make the proper diagnosis when encountering an incomprehensible clinical sign in the vicinity of the median nerve.