Infantile outcome at 3 years of age among monochorionic twins with Type-II or -III selective fetal growth restriction with isolated oligohydramnios who underwent fetoscopic laser photocoagulation.

OBJECTIVE: This study aimed to examine infantile outcomes at 3 years of age with selective fetal growth restriction (sFGR) Types II and III with isolated oligohydramnios who underwent fetoscopic laser photocoagulation (FLP). METHODS: This multicenter prospective cohort study included monochorionic diamniotic twins who underwent FLP for sFGR between 16 and 25 weeks of gestation. The indication for performing FLP was in cases of sFGR Type II or III with oligohydramnios, where the maximal vertical pocket was ≤2 cm among twins with FGR. This was done in the absence of a typical twin-twin transfusion syndrome diagnosis. The primary outcome was the intact survival (IS) rate of infants at the corrected age of 40 weeks and 3 years. IS at the corrected age of 40 weeks was defined as survival without grade III or IV intraventricular hemorrhage or cystic periventricular leukomalacia, and IS at 3 years of age was defined as survival without neurodevelopmental morbidity, including cerebral palsy, neurodevelopmental impairment with a total developmental quotient of ≤70, bilateral deafness, or bilateral blindness. RESULTS: Among 45 patients with sFGR, 30 (66.7%) were classified as having Type II and 15 (33.3%) as Type III sFGR. The prevalence of IS at the corrected age of 40 weeks was 51.1% (n=23) in FGR twins and 95.5% (n=42) in larger twins. The prevalence of IS at 3 years of age was 46.7% (n=21) in FGR twins and 86.4% (n=38) in larger twins. Among the 24 FGR twins who were not diagnosed with IS at 3 years of age, 91.7% (22 of 24 cases) suffered fetal or infantile demise other than miscarriage and neurodevelopmental impairment. All larger twins who were not diagnosed with IS at 3 years of age (n=6, 13.6%) had neurological morbidity, in addition to one case of miscarriage. CONCLUSIONS: FGR twins and larger twins, when subjected to FLP due to sFGR coupled with umbilical artery Doppler abnormalities and isolated oligohydramnios, exhibit low rates of neurological morbidity and low mortality, respectively. Therefore, FLP for Type II or III sFGR with oligohydramnios may be a feasible and preferable management option. This article is protected by copyright. All rights reserved.

Factors predictive of the presence of a CSF1R mutation in patients with leukoencephalopathy.

BACKGROUND AND PURPOSE: The purpose was to identify statistically factors that correlate with the presence of a colony-stimulating factor 1 receptor (CSF1R) mutation and to reevaluate the accuracy of the current diagnostic criteria for CSF1R-related leukoencephalopathy. METHODS: CSF1R testing was conducted on 145 consecutive leukoencephalopathy cases who were clinically suspected of having adult-onset leukoencephalopathy with axonal spheroids and pigmented glia. From these, 135 cases whose detailed clinical information was available were enrolled. Forward logistic stepwise regression was performed to generate a probability model to predict a positive CSF1R mutation result. The current diagnostic criteria were also applied to our cohort and their sensitivity and specificity were calculated. RESULTS: Twenty-eight CSF1R-mutation-positive cases and 107 CSF1R-mutation-negative cases were identified. Our probability model suggested that factors raising the probability of a CSF1R-mutation-positive result were younger onset, parkinsonism, thinning of the corpus callosum and diffusion-restricted lesions. It also showed that involuntary movements and brainstem or cerebellar atrophy were negative predictors of a CSF1R-mutation-positive result. In our cohort, the sensitivity and specificity for 'probable' or 'possible' CSF1R-related leukoencephalopathy were 81% and 14%, respectively. CONCLUSIONS: Clinical and brain imaging features predictive of the presence of a CSF1R mutation are proposed. Consideration of these factors will help prioritize patients for CSF1R testing.

Assessment of diastolic function of normal fetal heart using dual-gate Doppler.

OBJECTIVES: The ratio of mitral peak early diastolic filling to early diastolic mitral annular velocity (E/e') reflects diastolic cardiac function in adults. Dual-gate Doppler (DD) enables measurements of E/e' in the same heart beat. This study was designed to assess the utility of the DD method for measurement of fetal E/e' and determine reference ranges for normal fetuses. METHODS: This prospective study comprised normal singleton pregnancies undergoing fetal echocardiography between 16 and 36 weeks of gestation. According to the DD method, E-wave velocity on pulsed-wave Doppler and e'-wave on tissue Doppler imaging were measured simultaneously on an apical or basal four-chamber view, and fetal E/e' was calculated. Spearman's correlation coefficient was used to assess the relationship between gestational age (GA) and E-wave and e'-wave velocities and E/e'. RESULTS: A total of 133 pregnancies were included in this study and all E/e' measurements were successful. Significant correlation was observed between GA and both left ventricular (LV) E/e' (r s  = -0.666, P < 0.001) and right ventricular (RV) E/e' (r s  = -0.607, P < 0.001). The regression equations for bilateral E/e' were: LV-E/e' = 17.341 - 0.631GA + 0.008 × GA2 (mean ± SD, R 2  = 0.440 ± 1.333); and RV-E/e' = 19.156 - 0.794GA + 0.012GA2 (R 2  = 0.419 ± 1.329). CONCLUSIONS: Bilateral E/e' of normal fetuses, measured using the DD method, decreased with GA, which is considered to be related to myocardial maturity. DD is a useful and convenient method for evaluating fetal E/e' in order to assess diastolic function in the prenatal period. Copyright © 2017 ISUOG. Published by John Wiley & Sons Ltd.

Immunohistochemical analysis of gastrin-releasing peptide receptor (GRPR) and possible regulation by estrogen receptor ßcx in human prostate carcinoma.

Gastrin-releasing peptide (GRP) belongs to the family of bombesin-like peptides. GRP was demonstrated to stimulate the proliferation and invasiveness of androgen-independent prostate carcinoma. GRP mediates its action through the membrane-bound receptor, GRP receptor (GRPR), which is characterized by a high-affinity binding for both GRP and bombesin. In human prostate cancer tissue, GRPR mRNA was reported to be detectable in more than 90% but its immunolocalizaition has not been reported. Therefore, in this study we immunolocalized GRPR in 51 human prostate cancer cases and correlated the findings with several clinicopathological parameters in order to better understand the function and regulation of GRPR in human prostate cancer. GRPR was immnolocalized in carcinoma cells and their values were significantly associated with Gleason score and immunoreactivity of estrogen receptor ßcx (ERßcx) that is one of splicing variants of ligand dependent transcription factor, ERß, and considered to be prognostic factor of prostate cancer patients. The amounts of GRPR and ERßcx mRNA in three prostate cancer cell lines PC-3, DU-145 and LNCaP evaluated by quantitative RT-PCR (qPCR) analysis were also significantly correlated. In addition, we established stable transformants of prostate carcinoma cell line PC-3 introduced with ERßcx, and confirmed that GRPR mRNA was induced in ERßcx over-expressing PC-3 cells by qPCR analysis. These results also suggest that ERßcx contributes to prostate cancer development possibly through mediating GRPR expression in carcinoma cells.

Thermostable alanine dehydrogenase of Bacillus sp. DSM730: gene cloning, purification, and characterization.

We have cloned the thermostable alanine dehydrogenase (EC 1.4.1.1) gene from a thermophile, Bacillus sp. DSM730, into Escherichia coli C600 with a vector plasmid, pBR322. The enzyme was overproduced by the transformed cells, and purified to homogeneity with a yield of 69% by heat treatment and another step. The enzyme has a molecular weight of about 250,000 and consists of 6 subunits identical in molecular weight (43,000). It is not inactivated by heat treatment at 75 degrees C for 60 min, or incubation in the pH range of 5.5-10.5 at 55 degrees C for 10 min. The enzyme ctalyzes the oxidative deamination of L-serine in addition to L-alanine. The oxo analogue of serine is as reactive as pyruvate. Thus, the enzyme differs markedly from alanine dehydrogenases so far studied.

Stereospecific abstraction of epsilon-pro-R-hydrogen of L-lysine by L-lysine epsilon-dehydrogenase from Agrobacterium tumefaciens.

The stereochemical aspects of the L-lysine epsilon-dehydrogenase reaction were examined with (6R)-L-[6-3H]lysine and (6S)-DL-[6-3H]lysine. When (6S)-DL-[6-3H]lysine was used as a substrate, the tritium was found in the product, delta 1-piperideine-6-carboxylate. In contrast, the radioactivity from (6R)-L-[6-3H]lysine was not retained in the product. Thus, the pro-R hydrogen at the prochiral C-6 carbon of L-lysine is specifically abstracted by the enzyme: the enzyme behaves stereochemically as an amino acid D-dehydrogenase.

Activation of L-lysine epsilon-dehydrogenase from Agrobacterium tumefaciens by several amino acids and monocarboxylates.

The activation of lysine epsilon-dehydrogenase [EC 1.4.1.] by L-lysine was dependent on lysine concentration and was accompanied by association of the dimeric enzymes to a tetramer. The lysine concentration required for the half-maximal activation was 0.28 mM, which was lower than the Km value for L-lysine. In addition to L-lysine, several compounds, which were neither substrates nor inhibitors, activated the enzyme. The compounds which activated the enzyme have common structural characteristics: they have both a carboxyl group and a hydrophobic side chain. These activators also induced the association of the enzyme. The activation of the enzyme occurred well over the pH range 5.0 to 7.5, and the maximal activation was obtained by preincubation for 5 min at 30 degrees C and pH 7.4, when 5 mM L-lysine or 6-aminocaproate was used as an activator. NADH binding experiments indicated that about 2 mol of NADH bind to 1 mol of the tetrameric enzyme: the dimeric enzyme has one catalytic site. Binding experiments with n-[1-14C]heptanoate and L-[U-14C]lysine showed that approximately 2 mol of ligands bind to 1 mol of the dimeric enzyme and L-lysine could not bind to the catalytic site of the enzyme in the absence of NAD+. These results indicate the presence of one catalytic site and two activator binding binding sites in the dimeric enzyme.

An inducible NADP(+)-dependent D-phenylserine dehydrogenase from Pseudomonas syringae NK-15: purification and biochemical characterization.

An inducible NADP(+)-dependent D-phenylserine dehydrogenase [EC 1.1.1.-], which catalyzes the oxidation of the hydroxyl group of D-threo-beta-phenylserine, was purified to homogeneity from a crude extract of Pseudomonas syringae NK-15 isolated from soil. The enzyme consisted of two subunits identical in molecular weight (about 31,000). In addition to D-threo-beta-phenylserine, it utilized D-threo-beta-thienylserine, D-threo-beta-hydroxynorvaline, and D-threonine as substrates but was inert towards other isomers of beta-phenylserine and threonine. It showed maximal activity at pH 10.4 for the oxidation of D-threo-beta-phenylserine, and it required NADP+ as a natural coenzyme. NAD+ showed a slight coenzyme activity. The enzyme was inhibited by p-chloromercuribenzoate, HgCl2, and monoiodoacetate but not by the organic acids such as tartronate. The Michaelis constants for D-threo-beta-phenylserine and NADP+ were 0.44 mM and 29 microM, respectively. The N-terminal 27 amino acids sequence was determined. It suggested that the NADP(+)-binding site was located in the N-terminal region of the enzyme.

Properties of L-lysine epsilon-dehydrogenase from Agrobacterium tumefaciens.

Lysine epsilon-dehydrogenase, which has been purified to homogeneity from the extract of Agrobacterium tumefaciens ICR 1600, had a molecular weight of approximately 78,000 and consisted of two subunits identical in molecular weight (about 39,000). The enzyme showed a high substrate specificity. In addition to L-lysine, S-(beta-aminoethyl)-L-cysteine was deaminated by the enzyme, but to a far lesser extent. NAD+ and some NAD+ analogs (deamino-NAD+ and 3-acetylpyridine-NAD+) served as a cofactor. The pH optimum was at about 9.7 for the deamination of L-lysine. Although the NAD+ saturation curve was hyperbolic, a sigmoid saturation curve for L-lysine was obtained with the diluted enzyme solution, in which the dimeric enzyme was predominant. The reversible association of the enzyme to the tetramer was induced either by increasing the enzyme concentration or by addition of L-lysine. The preincubation of the enzyme with 5 mM L-lysine resulted in a 2-fold increase in the activity and gave a hyperbolic saturation curve for L-lysine. Upon modification of SH groups of the enzyme with DTNB, neither the interconversion between the dimer and the tetramer nor the activation by L-lysine occurred. These results indicated that the dimeric enzyme was activated by L-lysine and the activation resulted from the association of two dimeric enzymes to form a tetramer.

Recurrence in early gastric cancer.

In a retrospective study of 503 cases of early gastric cancer, 17 of the patients had died of a recurrence of the gastric cancer and 72 had died of unrelated causes. The cumulative recurrence mortality rates were 2.2% at 9 years for mucosal cancer and 8.4% at 8 years for submucosal cancer. The recurrence patterns of early gastric cancer were hematogenic metastasis to the liver, lung, or bone (nine cases), recurrence from lymph nodes (three cases), and recurrence in the residual stomach (five cases). Submucosal cancers with a macroscopically elevated appearance, lymph node metastasis, and evidence of vessel invasion were the high-risk cancers for hematogenic recurrence, and adjuvant chemotherapy should be prescribed. Two cases of lymph node recurrence were attributed to inadequacy of lymph node dissection. Because metastasis to the group 2 lymph nodes was noted in 1.5% of cases of early gastric cancer and a macroscopic diagnosis of nodal status was inaccurate, complete dissection should be performed regardless of identification of metastasis. Five cases of recurrence in the residual stomach were attributed to overlooked lesions of multiple carcinoma and were detected at an advanced stage. Careful and regular postoperative follow-up is required to detect these recurrences at an early stage.

Impairment of spatial memory and changes in astroglial responsiveness following loss of molar teeth in aged SAMP8 mice.

In order to evaluate the mechanism(s) responsible for senile impairment of cognitive function as a result of reduced mastication, the effects of the loss of the molar teeth (molarless condition) on the hippocampal expression of glial fibrous acidic protein (GFAP) and on spatial memory in young adult and aged SAMP8 mice were studied using immunohistochemical and behavioral techniques. Aged molarless mice showed a significantly reduced learning ability in a water maze test compared with age-matched control mice, while there was no difference between control and molarless young adult mice. Immunohistochemical analysis showed that the molarless condition enhanced the age-dependent increase in the density and hypertrophy of GFAP-labeled astrocytes in the CA1 region of the hippocampus. These effects increased the longer the molarless condition persisted. When the extracellular K+ concentration ([K+]o) was increased from 4 to 40 mM for hippocampal slices in vitro, the mean increase in the membrane potential was about 57 mV for fine, delicate astrocytes, the most frequently observed type of GFAP-positive cell in the young adult mice, and about 44 mV for the hypertrophic astrocytes of aged mice. However, there was no significant difference in resting membrane potential between these cell types. The data suggest that an impairment of spatial memory and changes in astroglial responsiveness occur following the loss of molar teeth in aged SAMP8 mice.

Lidocaine suppresses the sodium current in Euhadra neurons which is mediated by cAMP-dependent protein phosphorylation.

The action of a local anesthetic, lidocaine, in association with the cyclic AMP (cAMP)-mediated intracellular biochemical process, was examined in identified Euhadra neurons. Lidocaine dose-dependently inhibited the inward current which was elicited by dibutyryl cAMP (db-cAMP) and isobutylmethylxanthine (IBMX). This inhibitory effect was transiently reversed by the intracellular injection of a catalytic subunit of a cAMP-dependent protein kinase. The inward current elicited by db-cAMP and IBMX was abolished by Na(+)-free saline but not by Ca(2+)-free saline. The data suggest that lidocaine is not acting directly on the Na+ channel, but acts at a level proximal to the catalytic subunit of cAMP-dependent protein kinase.

Maturation of static sensitivity is related to expansion of the capsular space in rat buccal stretch receptors.

The developmental relationship between static sensitivity and structure of the buccal stretch receptor (BSR) in rats was investigated. When responses to ramp-and-hold stretches were recorded from isolated BSRs, their static sensitivities suddenly increased between 2 and 4 weeks after birth. However, no apparent change was observed throughout other developmental stages examined. Electron microscopic examination revealed a conspicuous expansion of the fluid-filled capsular space caused by completion of the outer capsule during the same postnatal period. These findings suggest that an increase in the capsular space of BSR may be involved in increasing the static sensitivity of this receptor.

Origin of primary sensory neurons innervating the buccal stretch receptor.

The primary sensory neurons innervating mechanoreceptors in oro-facial regions have their cell bodies in either the trigeminal ganglion or the mesencephalic nucleus of the trigeminal nerve. The buccal stretch receptor (BSR), a type of mechanoreceptor in the jaw of rodents, has recently been recognized as signaling the position of the mandible. The location of the primary afferent neurons innervating this receptor is unknown. To investigate the cell bodies of the BSR afferent neurons in rats, we applied wheat germ agglutinin-horseradish peroxidase (WGA-HRP) to the proximal stump of the severed nerve branch of the buccal nerve that supplied the BSR. HRP-labeled cell bodies were observed in the posterolateral portion of the ipsilateral trigeminal ganglion. None was found in the contralateral trigeminal ganglion or in the brainstem. All labeled cell bodies were oval or round and closely resembled pseudo-unipolar neurons. The mean diameter of the labeled somata ranged between 25.5 and 52.5 microm, with small (< or = 30 microm), medium (from 31 to 40 microm), and large somata (> or = 41 microm) accounting for 8.8%, 54.9%, and 36.3%, respectively. Among the myelinated nerve fibers in the branch in which WGA-HRP was applied, 78.5% terminated in the BSR and had larger fiber diameters than the rest, indicating that most of the medium and large HRP-labeled cell bodies were BSR afferents. From these results and the ontogenetic origin of this receptor, it is suggested that the BSR differentiated from the mechanoreceptors in the oral mucosa or the fascia of masticatory muscles.

The effect of cryotherapy on intraarticular temperature and postoperative care after anterior cruciate ligament reconstruction.

The objective of this study was to elucidate how cryotherapy after anterior cruciate ligament reconstruction affects intraarticular temperature and clinical results. A prospective and randomized study was performed on 21 knees of 21 patients. The ligament reconstruction was performed by single-incision arthroscopy using autogenous hamstring tendon. On completion of the surgery, thermosensors were implanted in the suprapatellar pouch and the intracondylar notch, and the intraarticular temperature was monitored while the joint was cooled. Cooling was performed in one group at 5 degrees C (N = 7) and in another at 10 degrees C (N = 7), for 48 hours. A control group (N = 7) did not undergo cryotherapy. The cooled groups showed three temperature phases: a low-temperature phase immediately after the ligament reconstruction, followed by a temperature-rising phase and a thermostatic phase. The control group had no low-temperature phase and immediately entered a thermostatic phase. During the low-temperature phase in the treated groups, the temperature of the suprapatellar pouch and of the intercondylar notch were significantly lower than the body temperature. The pain score and the number of times an analgesic had to be administered were both significantly lower in the 10 degrees C group than in the control group. Blood loss was significantly less in the 5 degrees C group than in the control group.

Epidermal growth factor in gastric carcinoma as a risk factor of postoperative recurrence.

The expression of epidermal growth factor (EGF) was immuno-histochemically examined in the primary lesions of 201 patients with advanced gastric cancer who underwent curative resection. EGF-positive tumor cells were detected in the specimens of 75 patients (37.3%). A significant correlation was observed between the extent of lymph node metastasis and EGF expression (p < 0.01), indicating that EGF-positive cancer has a tendency to lymph node metastasis. The 2-year and 5-year survival rates of the patients with EGF-negative tumors were 77.2% and 63.0%, respectively, while those with EGF-positive tumors were 59.1% and 47.5% respectively. The difference in survival rates reached a maximum 2 years after surgery, indicating that the patients with EGF-positive gastric cancer had a higher risk of early recurrence. Therefore, the amount of EGF produced by a tumor may play some role in the growth of metastatic tumors and residual tumor cells after surgery, and thus be a potential risk factor in recurrence.

Surgical approaches in primary gastric lymphoma and carcinoma.

Data on 29 patients with primary gastric lymphoma and on 1655 with gastric carcinoma were compared with respect to the local extension and the results of surgery. More than two-thirds of the gastric area was involved due to multiple or widespread lesion in 25.9% and 7.0% of the patients with gastric lymphoma and carcinoma, respectively. Maximum tumor dimension exceeded 10 cm in 33.3% and 9.5% of gastric lymphomas and carcinomas respectively. Involvement of the extra-perigastric lymph nodes was evident in 51.8% and 26.2% of the patients with gastric lymphoma and carcinoma respectively. Therefore, wide and extended resection, including total gastrectomy, would be required more often for the surgical excision of gastric lymphoma. Peritoneal dissemination and liver metastasis were much less common in those patients with gastric lymphoma and hence, benefiting most from surgery. The five year survival rate after curative resection was somewhat better in patients with a gastric lymphoma (79.3% as compared to 65.6%), whereas the prognosis after non-curative surgery was equally unfavorable. Aggressive surgical excision is thus recommended for the treatment of patients with a primary gastric lymphoma.

Recurrent pattern of digestive tract carcinoma in the Japanese: comparison of gastric cancer to colon cancer.

Six hundred and two consecutive cases of gastric cancer and 204 of colorectal cancer in recent decades were investigated for recurrence. Recurrence occurred in 103 cases of gastric cancer and 31 cases of colorectal cancer. We classified three categories of recurrent patterns; A: local, B: peritoneal dissemination, and C: distant metastasis. In gastric cancer, 11 cases (11%) were grouped A, 50 (48%) were B, and 42 (41%) were C. Similarly, 12 (39%) were grouped A, 5 (16%) were B, and 14 (45%) were C in colorectal cancer. The incidence of local recurrence was more increased in colorectal cancer than in gastric cancer (p < 0.01). On the contrary, that of peritoneal dissemination was more increased in gastric cancer than in colorectal cancer (p < 0.01). In gastric cancer, 8 cases of 103 survived more than 5 years after operation, furthermore 7 cases obtained more than 5 years of tumor-free interval. At the time of diagnosis of recurrence, data of serum CEA was available in 77 for gastric cancer and 29 for colon cancer. CEA positive cases were revealed 32 (42%) for gastric cancer and 24 (83%) for colorectal cancer (p < 0.01). Our data clarified the different recurrence patterns between gastric cancer and colorectal cancer. And suggest that more than 5 years follow-up should be needed in patients with gastric cancer.

Intra-operative assessment of lymph node metastasis for colorectal carcinoma.

We have evaluated the reliability of intra-operative diagnosis for lymph node metastases and assessed the clinical features affecting accuracy of the intra-operative diagnosis based on a review of operative and pathological records in 218 patients treated by curative surgery for colorectal carcinoma. The sensitivity and specificity for diagnosis of lymph node metastases was 74.2% and 54.5%, respectively. The clinically positive but pathologically negative [cN(+),pN(-)] lymph nodes group differed significantly from the cN(-),pN(-) group with respect to tumor size, gross appearance and depth of tumor invasion. In the cN(+),pN(+) group, the tumor was larger and depth of tumor invasion was more extensive than in the cN(-),pN(+) group. The postoperative survival curves showed a statistically significant difference between cN(-),pN(-) and cN(+),pN(-) groups (P<0.05). Our data suggest less than satisfactory results for the intra-operative diagnosis of lymph node metastases. Macroscopically, tumor size and gross appearance were important clinical characteristics affecting accuracy in the macroscopic diagnosis of lymph node metastases. Therefore, prophylactic lymph node dissection is recommended, regardless of intra-operative assessment of lymph node metastases.