Benign hilar bile duct strictures resected as perihilar cholangiocarcinoma.

BACKGROUND: Differentiation between perihilar cholangiocarcinoma (PHCC) and benign strictures is frequently difficult. The aim of this study was to investigate the incidence and long-term outcome of patients with tumours resected because of suspicion of PHCC, which ultimately turned out to be benign (malignancy masquerade). METHODS: Patients who underwent surgical resection with a diagnosis of PHCC between 2001 and 2016 were reviewed retrospectively. RESULTS: Among 707 consecutive patients, 685 had PHCC and the remaining 22 (3·1 per cent) had benign biliary stricture. All patients with benign disease underwent major hepatectomy, with no deaths. Preoperative histological assessment using bile duct biopsy or aspiration cytology had a high specificity (90 per cent), low sensitivity (62 per cent) and unsatisfactory accuracy (63 per cent). Despite the increasing use of histological assessment, the incidence of benign strictures resected did not decrease over time, being 0·9 per cent in 2001-2004, 4·0 per cent in 2005-2008, 3·8 per cent in 2009-2012 and 2·9 per cent in 2013-2016. The final pathology of benign strictures included IgG4-related sclerosing cholangitis (9 patients), hepatolithiasis (4), granulomatous cholangitis (3), non-specific chronic cholangitis (3), benign strictures after cholecystectomy (2), and a benign stricture possibly caused by parasitic infection (1). The 10-year overall survival rate for the 22 patients with benign stricture was 87 per cent, without recurrence of biliary stricture. CONCLUSION: The incidence of benign strictures resected as PHCC as a proportion of all resections was relatively low, at 3·1 per cent. Currently, unnecessary surgery for suspected PHCC is unavoidable.

ANTECEDENTES: La diferenciación entre colangiocarcinoma perihilar (perihilar colangiocarcinoma, PHCC) y estenosis benignas es con frecuencia difícil. El objetivo de este estudio fue investigar la incidencia y el resultado a largo plazo de los tumores resecados con sospecha diagnóstica de PHCC, que finalmente resultaron ser benignos (malignidad enmascarada). MÉTODOS: Se revisaron retrospectivamente los pacientes con diagnóstico de PHCC que se sometieron a resección quirúrgica entre 2001 y 2016. RESULTADOS: Entre 707 pacientes consecutivos, 685 pacientes presentaban PHCC y los 22 restantes (3,1%) tenían una estenosis biliar benigna. Todos los pacientes con patología benigna se sometieron a una hepatectomía mayor, sin mortalidad. La evaluación histológica preoperatoria mediante biopsia de conducto biliar o citología por aspiración tuvo una alta especificidad (90%), una baja sensibilidad (62%) y una exactitud diagnóstica insatisfactoria (63%). A pesar del uso creciente de la evaluación histológica, la incidencia de estenosis benignas resecadas no disminuyó con el tiempo, con un 0,9% en 2001-2004, un 4,0% en 2005-2008, un 3,8% en 2009-2012 y un 2,9% en 2013-2016. La patología final de las estenosis benignas incluyó colangitis esclerosante relacionada con IgG4 (n = 9), hepatolitiasis (n = 4), colangitis granulomatosa (n = 3), colangitis crónica no específica (n = 3), estenosis benignas tras una colecistectomía (n = 2) y una estenosis benigna posiblemente causada por una infección parasitaria (n = 1). Los resultados a largo plazo de los 22 pacientes con estenosis benigna fueron mejores (tasa de supervivencia a 10 años; 87,4%) sin recidiva de la estenosis biliar. CONCLUSIÓN: La incidencia de pacientes con estenosis benignas resecadas como PHCC en comparación con todas las resecciones fue relativamente baja, del 3,1%. Actualmente, la cirugía "innecesaria" por sospecha de PHCC es inevitable.

Major hepatectomy with or without pancreatoduodenectomy for advanced gallbladder cancer.

BACKGROUND: The indications for major hepatectomy for gallbladder cancer either with or without pancreatoduodenectomy remain controversial. The clinical value of these extended procedures was evaluated in this study. METHODS: Patients who underwent major hepatectomy for gallbladder cancer between 1996 and 2016 were identified from a prospectively compiled database. Postoperative outcomes and overall survival were compared between patients undergoing major hepatectomy alone or combined with pancreatoduodenectomy (HPD). RESULTS: Seventy-nine patients underwent major hepatectomy alone and 38 patients had HPD. The patients who underwent HPD were more likely to have T4 disease (P < 0·001), nodal metastasis (P = 0·015) and periaortic nodal metastasis (P = 0·006), but were less likely to receive adjuvant therapy (P = 0·006). HPD was associated with a high incidence of grade III or higher complications (P = 0·002) and death (P = 0·037). Overall survival was longer in patients who underwent major hepatectomy alone than in patients who underwent HPD (median survival time 32 versus 10 months; P < 0·001). In multivariable analysis, surgery in the early period (1996-2006) (P = 0·002), pathological T4 disease (P = 0·005) and distant metastasis (P < 0·001) were associated with shorter overall survival, and cystic duct tumour (P = 0·002) with longer overall survival. CONCLUSION: Major hepatectomy alone for gallbladder cancer contributes to favourable overall survival with low morbidity and mortality, whereas HPD is associated with poor overall survival and high morbidity and mortality rates. HPD may eradicate locally spreading gallbladder cancer; however, the indication for the procedure is questioned from an oncological viewpoint.

Ethanol ablation for refractory bile leakage after complex hepatectomy.

BACKGROUND: Only a few reports exist on the use of ethanol ablation for posthepatectomy bile leakage. The aim of this study was to assess the value of ethanol ablation in refractory bile leakage. METHODS: Medical records of consecutive patients who underwent a first hepatobiliary resection with bilioenteric anastomosis between 2007 and 2016 were reviewed retrospectively, with special attention to bile leakage and ethanol ablation therapy. Bile leakage was graded as A/B1/B2 according to the International Study Group of Liver Surgery definition. Absolute ethanol was injected into the target bile duct during fistulography. RESULTS: Of the 609 study patients, 237 (38·9 per cent) had bile leakage, including grade A in 33, grade B1 in 18 and grade B2 in 186. Left trisectionectomy was more often associated with grade B2 bile leakage than other types of hepatectomy (P < 0·001). Of 186 patients with grade B2 bile leakage, 31 underwent ethanol ablation therapy. Ethanol ablation was started a median of 34 (range 15-122) days after hepatectomy. The median number of treatments was 3 (1-7), and the total amount of ethanol used was 15 (3-71) ml. Complications related to ethanol ablation included transient fever (27 patients) and mild pain (13). Following ethanol ablation, bile leakage resolved in all patients and drains were removed. The median interval between the first ablation and drain removal was 28 (1-154) days. CONCLUSION: Ethanol ablation is safe and effective, and may be a treatment option for refractory bile leakage.

Use of loop-mediated isothermal amplification of the IS900 sequence for rapid detection of cultured Mycobacterium avium subsp. paratuberculosis.

We evaluated the usefulness of loop-mediated isothermal amplification (LAMP) in detecting specific gene sequences of Mycobacterium avium subsp. paratuberculosis (MAP). A total of 102 primer sets for LAMP was designed to amplify the IS900, HspX, and F57 gene sequences of MAP. Using each of two primer sets (P-1 and P-2) derived from the IS900 fragment, it was possible to detect MAP in a manner similar to that used with nested PCR. The sensitivity of LAMP with P-1 was 0.5 pg/tube, which was more sensitive than nested PCR. When P-2 was used, 5 pg/tube could be detected, which was the same level of sensitivity as that for nested PCR. LAMP with P-1 was specific. Although only 2 Mycobacterium scrofulaceum strains out of 43 non-MAP mycobacterial strains were amplified, the amplification reaction for these strains was less efficient than for MAP strains, and their products could be distinguished from MAP products by restriction digestion. LAMP with P-2 resulted in very specific amplification only from MAP, the same result obtained with nested PCR. Our LAMP method was highly specific, and the white turbidity of magnesium pyrophosphate, a by-product of the LAMP reaction, allowed simple visual detection. Our method is rapid, taking only 2 h, compared with 4 h for nested PCR. In addition, the LAMP method is performed under isothermal conditions and no special apparatus is needed, which makes it more economical and practical than nested PCR or real-time PCR. These results indicate that LAMP can provide a rapid yet simple test for the detection of MAP.

Botryoid Wilms tumor: case report and review of literature.

A rare case of botryoid Wilms tumor is presented. The main clinical manifestations were persistent low-grade fever, malaise, and proteinuria associated with microhematuria. Ultrasonography revealed an echogenic mass in the right kidney, and a contrast-enhanced mass was found in the dilated collecting system by contrast-enhanced computed tomography. The surgically resected tumor was a polypoid, light-yellow, glistening mass that occupied a large part of the renal pelvis and originated from the pelvicaliceal wall. Part of the tumor extended to the proximal ureter, resulting in hydronephrosis in the involved kidney. No parenchymal lesion was observed. Microscopic examination revealed epithelial, stromal, and blastemal components, which indicated Wilms tumor. Infection had occurred in the hydronephrotic kidney, which presumably had caused the major presenting symptoms. The prognosis of our patient and previously reported cases of botryoid Wilms tumor was good compared with that of typical Wilms tumor, since the botryoid type can be detected at an early stage.

Rapid analysis of allelic variants of the sheep PrP gene by oligonucleotide probes.

A rapid method to determine the allelic variants of the sheep PrP gene was developed. DNA samples from 128 Suffolk sheep (39 rams and 89 ewes) were screened by using polymerase chain reactions and dot-blot hybridization with 32P-labeled nine allele-specific oligonucleotide probes corresponding to the polymorphic PrP codons 112, 136, 154 and 171. Three allelic variants of the PrP gene, PrP(MARQ), PrP(TARQ) and PrP(MARR), were found in the flocks. Among those variants, nearly half of the ewes had alleles of the 171-Arg variant that is closely associated with resistance to natural scrapie. Assessments of allelic mutations of the PrP gene may help to select the scrapie-resistant progenitors in the flocks.

[Ocular complications after renal transplantation].

Seventy-two post-renal transplant patients were studied for ocular complications. Of 72 patients, 56 (77.8%) showed some ocular abnormality. Steroid cataract was the most common complication, occurring in 45 patients (62.5%). Eleven patients (18 eyes) had undergone operations for cataract. The average of their ages was 39.7 years and the period from renal transplantation to cataract operation was 3.3 years. Postoperative visual acuity was over 20/20 in most cases. Increased intraocular pressure was encountered in 9 patients (12.5%), cytomegalovirus ocular infection in 2 (2.8%), hypertensive retinopathy in 2 (2.8%), branch retinal vein occlusion in 1 (1.4%), and subconjunctival hemorrhage in 15 (20.8%). A new immunosuppressant, cyclosporine, increased renal graft survival more than azathioprine. However, ocular complications such as steroid cataract occurred frequently in spite of the use of cyclosporine, as in the azathioprine era. In conclusion, it is necessary for renal transplant patients to receive a periodical ophthalmological check-up.

Vasomotor nerves of vessels in the human optic nerve.

Aminergic and cholinergic vasomotor nerves in vessels of the human optic nerve were studied morphologically. Aminergic nerve fibers were observed by the glyoxylic acid method. Cholinergic nerve fibers were observed by light microscopy after acetylcholinesterase staining by the Karnovsky-Roots method and Tago's modified method. In the retrobulbar optic nerve behind the bulbus, aminergic and cholinergic vasomotor nerves were observed to be dense in the central retinal artery and vein and posterior ciliary arteries. A large number of vasomotor nerves were also demonstrated in vessels in the septum of the optic nerve, but they were sparse in pial vessels. Further centrally, a few vasomotor nerves were found in pial vessels of the intracanalicular and intracranial optic nerve, but few were observed in the septum of the optic nerve. At the optic chiasm they were densely distributed in pial vessels.

Antigenic alteration in major piroplasm surface proteins of Theileria sergenti during infection.

Theileria sergenti piroplasms were purified from different parasitemia peaks of cattle infected with parasitized erythrocytes or sporozoites during persistent infection. Their reactivities with monoclonal antibodies 13F5 and C9, which recognize 23 kDa and 32 kDa piroplasm surface proteins, respectively, were analyzed. Antigenic differences were observed among parasites from different parasitemia peaks during persistent infection when cattle were infected with sporozoites. Results of two-dimensional polyacrylamide gel electrophoresis showed that the 23 and 32 kDa proteins were expressed in all samples tested, regardless of their reactivities with the monoclonal antibodies. In contrast, parasites obtained from cattle inoculated with parasitized erythrocytes showed no antigenic alteration over a 2 month observation period. The results suggest that antigenic alteration of T. sergenti during persistent infection is related to whether the parasites proliferate through extraerythrocytic schizont stage in cattle or sporozoite and other sexual stages in tick vector.

Cloning of complementary DNA encoding a 135-kilodalton protein secreted from porcine corpus epididymis and its identification as an epididymis-specific alpha-mannosidase.

In the preceding study (Okamura et al., 1992; Biol Reprod 47:1040-1052) we suggested that a 135-kDa protein secreted by porcine epididymis is involved in the sperm maturation. In this work, we have isolated the cDNA clone coding the 135-kDa protein in an effort to investigate its structure and function. The 135-kDa protein was purified from porcine cauda epididymal fluid. Three oligonucleotide probes were synthesized according to the amino acid sequences of N-termini of the native protein and trypsin-digested peptides. A cDNA clone hybridizing with these three probes was isolated from the cDNA library derived from the porcine proximal corpus epididymis. It encodes a novel protein with 1,006 amino acid residues in an open reading frame. Its overall amino acid sequence was significantly homologous (25.7%) to the alpha-mannosidase precursor of Dictiostelium discoideum (P34098). The 135-kDa protein could digest both p-nitro-phenyl-alpha-D-mannoside and high mannose oligo saccharide (Man8-GlcNAc2), strongly suggesting that it is an alpha-mannosidase homologue. The expression of this protein was specific to porcine and was localized to the very narrow parts of epididymis: the border of the caput and corpus epididymis. This protein may serve as a good marker for the functional differentiation in porcine epididymis. A possible role of this protein in the species-specific sperm-egg interaction is discussed.

Molecular mechanisms of the activation of maturation-promoting factor during goldfish oocyte maturation.

Oocyte maturation is triggered by the activation in the oocyte cytoplasm of maturation-promoting factor (MPF), which consists of cdc2 (a catalytic subunit) and cyclin B (a regulatory subunit). Immature goldfish oocytes contain only inactive monomeric 35-kDa cdc2 and do not stockpile cyclin B. In maturing oocytes, activation of cdc2 is associated with its Thr161 phosphorylation and mobility shift on SDS-PAGE from 35 to 34 kDa after binding to cyclin B. Using mutant cdc2, we show that Thr161 phosphorylation is required for both the downward shift and the kinase activation. Since cdc2 Tyr15 is not phosphorylated after binding to cyclin B, it does not require dephosphorylation. This situation is obviously different from that in immature Xenopus oocytes, in which the cdc2-cyclin B complex preexists with cdc2 phosphorylated on both Tyr15 and Thr161, thereby requiring Tyr15 dephosphorylation catalyzed by cdc25 phosphatase for MPF activation. These results indicate that these species employ different mechanisms of MPF activation during oocyte maturation, although the final molecular structure of the active MPF (cdc2 bound to cyclin B and phosphorylated on Thr161) is identical.

Gangliosides as a possible receptor on the bovine erythrocytes for Theileria sergenti.

To elucidate whether or not gangliosides on the bovine erythrocytes serve as a receptor for Theileria sergenti merozoites, the reactivities of the T. sergenti piroplasms with gangliosides were studied by the liposome agglutination test. The parasites reacted weakly with I-active ganglioside containing N-acetylneuraminic acid (NeuAc) and strongly with I-active ganglioside containing N-glycolylneuraminic acid (NeuGc). However, none of the other gangliosides expressed on the bovine erythrocytes, such as GM3 (NeuAc), GM3 (NeuGc), sialosylparagloboside (SPG) (NeuAc), SPG (NeuGc), i-active ganglioside (NeuAc), and i-active ganglioside (NeuGc), were recognized. After infection with T. sergenti, furthermore, the content of I-active ganglioside (NeuAc) was less (p < 0.05), and I-active ganglioside (NeuGc) content was much less in the erythrocytes (p < 0.01), though the contents of other NeuAc- and NeuGc-containing gangliosides did not so vary with T. sergenti infection. These results suggest that the parasites recognize the I-active ganglioside as their receptor and bind preferentially to NeuGc-carrying I-active ganglioside rather than to NeuAc-type in the target cell membranes, and that the reduction of the contents of I-active gangliosides (NeuAc and NeuGc) on the erythrocytes was related to T. sergenti infection.

Alteration of ganglioside composition in the erythrocytes associated with Theileria sergenti infection.

The changes in ganglioside composition of bovine erythrocytes associated with Theileria sergenti infection were investigated using the erythrocytes before and after the infection. The erythrocytes before infection with T. sergenti had GM3, sialosylparagloboside (SPG), i-active, and I-active ganglioside as predominant gangliosides. After infection with T. sergenti merozoites, the contents of SPG and i-active ganglioside were slightly less, and I-active ganglioside content was much less in the erythrocytes, though GM3 content did not so vary. The decreased I-active ganglioside content showed a recovery as the parasitemia waned to low level in the infected cattle. The total amount of lipid-bound sialic acid also decreased in the erythrocytes after the infection. Similar changes were also caused by the incubation of liposomes containing ganglioside fraction obtained from bovine erythrocytes with T. sergenti piroplasms. These results suggest that the reduction of the contents of SPG, i-active, and I-active ganglioside on the erythrocytes was related to the T. sergenti infection.

Delayed myelination in a patient with 18q- syndrome.

A Japanese boy with the typical manifestations of 18q-syndrome and delayed myelination on magnetic resonance imaging is described. Cytogenetic investigation revealed a deletion at 18q21.3. Three serial magnetic resonance images demonstrated that myelination in the central nervous system was delayed except for the corpus callosum and brainstem. This pattern of delayed myelination appears to be peculiar to the 18q- syndrome. Because the gene for myelin basic protein has been localized to the distal end of the long arm of chromosome 18, we speculate that the abnormal myelination in our patient was partly due to the failure of expression of the myelin basic protein gene.

Detection of Theileria sergenti infection in cattle by polymerase chain reaction amplification of parasite-specific DNA.

A pair of synthetic oligonucleotide primers, designed from the gene encoding a 32-kDa intraerythrocytic piroplasm surface protein of Theileria sergenti, were used to amplify parasite DNA from the blood of T. sergenti-infected cattle by means of the polymerase chain reaction (PCR). PCR-amplified DNA was examined by electrophoresis and by dot blot or microplate hybridization using a parasite-specific cDNA probe. PCR was specific for T. sergenti, since no amplification was detected with DNA from Anaplasma centrale, Babesia ovata, uninfected erythrocytes, and leukocytes. This method was sensitive enough to detect about 4.5 parasites per microliters of blood with a 10-microliters sample volume. Moreover, of 66 specimens from grazing cattle, 40 were microscopically positive, whereas PCR revealed that 54 samples were positive. Therefore, PCR provides a useful diagnostic tool for detecting T. sergenti-infected cattle, and it is significantly more sensitive than the current methods.

Changes in the hybridization patterns of populations of Theileria sergenti during infection.

Restriction fragment length polymorphisms (RFLPs) of Theileria sergenti DNA were analysed using probes of a genomic DNA fragment (pTs 2) and a cDNA corresponding to this genomic probe (C-Ts 2). Each of the probes detected RFLPs in DNA from different stocks of Theileria sergenti. Additionally, using these probes, alterations in hybridization patterns were observed in samples of the parasites harvested at different times after individual calves had been infected with Theileria sergenti. This result suggests that the Theileria sergenti stocks used were mixed parasite populations.

Spectral sensitivity of monochromatic ERG c-wave of chicken under color adaptation.

The spectral sensitivity of the ERG c-wave was studied in the chicken under yellow or red light adaptation of various intensities, ie, 1, 3, 4 and 5 W/m2 at the corneal surface in the former, and 3, 4 and 5 W/m2 in the latter. The peak wavelength of the spectral sensitivity curve of the c-wave amplitude under both yellow and red light adaptations was 520 or 540 nm, being shorter as compared with the peak wavelength (560 nm) under white light adaptation (Fukuda, 1989). Under the yellow or red light adaptation the sensitivity of the c-wave was suppressed at a range of longer wavelength, suggesting a possible isolation of the blue and green cone-driven c-wave responses. The peak (520 or 540 nm) and the shoulder (580 nm) in the spectral sensitivity curves were presumed to be derived from the three cone systems.

Adult T-cell leukemia with leukemic cell infiltration in the conjunctiva. A case report.

A 77-year-old Japanese woman who had suffered from skin eruptions since 1986 was admitted in January, 1990. A diagnosis of adult T-cell leukemia/lymphoma (ATL) was made on the basis of clinical and laboratory data. On admission, erythematous lesions were present in both eyelids. Yellowish-white elevated lesions were found along the limbal conjunctiva, and extended segmentally into the cornea of both eyes. Microscopically, leukemic infiltration into the subepithelial layer of the conjunctiva was observed. Ophthalmic manifestations in ATL have not been well described, because of a little attention paid to them.

Localization of a maturation-dependent epididymal sperm surface antigen recognized by a monoclonal antibody raised against a 135-kilodalton protein in porcine epididymal fluid.

A specific 135-kDa protein was purified from porcine cauda epididymal fluid. Analysis of its N-terminal amino acid sequence revealed it to be a new protein. Stable clones of hybridomas that produced monoclonal antibodies against the purified 135-kDa protein were established. A clone, B-11, reacting both with epididymal fluid and with sperm plasma membranes was selected and used in this study. Immunoblotting analysis showed that B-11 reacted only with a 135-kDa protein among epididymal fluid proteins. In contrast, B-11 did not recognize a similar 135-kDa sperm protein but did strongly react with a 27-kDa protein among sperm membrane proteins, extracted by NP-40 in the presence of protease inhibitors. B-11 also reacted only with a 27-kDa protein fragment among trypsin digests of the 135-kDa epididymal protein. The 135-kDa protein was first detected, by ELISA or immunoblotting analysis, at the beginning of the corpus epididymis. Maximal levels were reached in the distal corpus and levels were slightly decreased in the cauda epididymis. On the other hand, the surface of caput sperm were found to contain small amounts of antigen(s), the concentration of which gradually increased during epididymal transit. In immunocytochemical studies, the antigen was detectable in the epithelial cells from the initial segment to the corpus of the epididymis but not in the caudal cells. In the lumen, the presence of the 135 kDa protein was apparent in the corpus (at a maximum in the middle and distal corpus) and to a lesser degree in the caudal lumen. The 27-kDa protein was distributed all over the equatorial region of the acrosome of less than 10% of caput epididymal sperm. As sperm passed through the corpus epididymis, the percentage of immunoreactive cells increased and the protein was restricted to specific domains of the sperm head. Thus, on the mature sperm, antigen was localized in a crescent-shaped area of the equatorial segment just behind the anterior part of the acrosome and on the apical rim of the sperm head. This is the first observation of a sperm surface antigen derived from an epididymal protein as a proteolytic fragment that interacts with specific regions of the sperm membrane during the process of spermatozoa maturation.