Amplified polarization properties of electrospun nanofibers containing fluorescent dyes and helical polymer.

Nanofibers with the cationic fluorescent molecules, 1-pyrenemethylamine hydrochloride (PyrMA) and α,ß,γ,δ-tetrakis(1-methylpyridinium-4-yl)porphyrin p-toluenesulfonate (TMPyP), in the presence of complementary anionic chiral polysaccharide, hyaluronic acid (HA) and transparent polymer, polyethylene oxide (PEO) were successfully prepared using an electrospinning technique. The results showed that the fluorescence spectra of electrospun nanofibers (ENFs) of PEO-HA-PyrMA exhibit a higher amount of pyrene excimer emission when the [PyrMA]/[HA] ratio was increased from 0.5 to 1, while the concentration of TMPyP had no remarkable effect on the fluorescence of the PEO-HA-TMPyP ENFs. From circular dichroism (CD) characterization, the ENFs of both dyes exhibit stronger intensities in the apparent CD spectra upon increasing the dye ratio. Moreover, they also showed higher CD intensities when compared with those of solutions and thin films prepared using other methods. The apparent CD in this work was proved to mainly originate from linear dichroism (LD) and linear birefringence (LB) obtained by macroscopic alignment along the electrospinning process. These results suggest the advantages of molecular orientation via electrospinning, together with chirality induction by incorporation of a chiral HA polymer. The enhancement of these polarization properties provides great possibilities for improving fluorescent-chiral nanofibers for further optoelectronic applications.

Specific excitonic interactions in the aggregates of hyaluronic acid and cyanine dyes with different lengths of methine group.

The absorption and circular dichroism (CD) spectra of three types of cyanine dyes with different lengths of methine group (3,3'-diethylthiadicarbocyanine iodide, DTDC; 3,3'-diethylthiacarbocyanine iodide, DTC; and 3,3'-diethylthiacyanine iodide, DTTHC) in an aqueous solution were compared with and without hyaluronic acid (HA), which has a helical structure. DTDC forms chiral H- and J-aggregates, whereas DTC and DTTHC are unable to form any aggregates. DTDC also forms H- and J-aggregates in the presence of sodium polyacrylate (PA) with a random-coil structure; however, the PA-DTDC aggregates exhibit no chirality. These results suggest that the chirality of HA-DTDC aggregates is induced by the helical structure of HA. In 2.4 vol% and 10 vol% methanol, HA-DTDC aggregates displayed different patterns of temperature dependence, whereas no aggregation was observed in 30 vol% methanol. The solubility of DTDC in a mixed solvent of water and methanol is generally improved by the addition of methanol, which prevents the aggregation of DTDC.

Faecal lactoferrin is a useful biomarker for mucosal healing in patients with ulcerative colitis during granulocyte and monocyte adsorptive apheresis therapy.

AIM: The study investigated the value of faecal lactoferrin as a follow-up biomarker for mucosal healing of ulcerative colitis during granulocyte and monocyte adsorptive apheresis (GMA) therapy. METHOD: Patients with ulcerative colitis exhibiting a moderate or severe disease activity with a partial Mayo Score (pMS) of over 4 were enrolled in this study. The patients received 10 courses of GMA therapy. The pMS value and faecal lactoferrin level were monitored and compared with the findings of endoscopy until 12 months after the last dose of GMA therapy. RESULTS: Twenty patients (male:female 11:9) were enrolled in this study. Twelve had total colitis, while six had left-sided involvement and two had distal proctitis. Thirteen (65.0%) responded to GMA therapy. The faecal lactoferrin levels were significantly decreased in patients who responded to GMA therapy (P < 0.05), whereas the levels did not change in non-responders. Moreover, the faecal lactoferrin levels correlated with the endoscopic findings (r = 0.792, P < 0.01) and pMS scores (r = 0.529, P < 0.01). The correlation coefficients between the faecal lactoferrin levels and mucosal findings were higher than those observed between the pMS score and mucosal findings. CONCLUSION: The faecal lactoferrin level is a useful biomarker of the mucosal findings in ulcerative colitis. Although endoscopy is the gold standard, the faecal lactoferrin level can be used as a biomarker during GMA therapy in patients with ulcerative colitis.

Experimental observation of the role of mutual information in the nonequilibrium dynamics of a Maxwell demon.

We validate experimentally a fluctuation relation known as generalized Jarzynski equality governing the work distribution in a feedback-controlled system. The feedback control is performed on a single electron box analogously to the original Szilard engine. In the generalized Jarzynski equality, mutual information is treated on an equal footing with the thermodynamic work. Our measurements provide the first evidence of the role of mutual information in the fluctuation theorem and thermodynamics of irreversible processes.

Ramucirumab plus FOLFIRI as second-line treatment for patients with RAS wild-type metastatic colorectal cancer previously treated with anti-EGFR antibody: JACCRO CC-16.

BACKGROUND: Chemotherapy in combination with anti-epidermal growth factor receptor (EGFR) antibody is considered a first-line treatment regimen for RAS wild-type and left-sided metastatic colorectal cancer (mCRC), whereas second-line treatment regimens have not yet been established. Few studies have prospectively evaluated second-line treatment with anti-vascular endothelial growth factor antibody after first-line anti-EGFR antibody therapy for RAS wild-type mCRC. PATIENTS AND METHODS: This non-randomized phase II trial investigated the clinical outcomes of second-line ramucirumab (RAM) plus fluorouracil, levofolinate, and irinotecan (FOLFIRI) after first-line anti-EGFR antibody in combination with doublet or triplet regimen in patients with RAS wild-type mCRC. The primary endpoint was the 6-month progression-free survival (PFS) rate. The secondary endpoints were PFS, overall survival (OS), objective response rate (ORR), rate of early tumor shrinkage (ETS), and safety. We hypothesized a threshold 6-month PFS rate of 30% and an expected 6-month PFS rate of 45%. Treatment was considered effective if the lower limit of the 90% confidence interval (CI) of the 6-month PFS rate was >0.30. RESULTS: Ninety-two patients were enrolled in the study. The primary tumor was located on the left side in 86 (95.6%) patients. Twenty (22.0%) patients had received triplet plus cetuximab as previous therapy. Six-month PFS rate was 58.2% (90% CI 49.3% to 66.2%) with a median PFS of 7.0 months (95% CI 5.7-7.6 months). Median OS was 23.6 months (95% CI 16.5-26.3 months). The ORR and ETS rate were 10.7% and 16.9%, respectively, in 83 patients with measurable lesions. The 6-month PFS rate was comparable between patients previously treated with doublet and triplet regimens; however, median PFS was longer for the doublet regimen (7.4 versus 6.4 months, P = 0.036). CONCLUSIONS: Our study demonstrated prospectively that RAM plus FOLFIRI is an effective second-line treatment after anti-EGFR antibody-containing first-line therapy in RAS wild-type and left-sided mCRC. Furthermore, the results were similar for patients who were previously treated with triplet regimen.

The Culdotomy FourS Two U procedure for transvaginal access to the peritoneal cavity.

A reliable method of transvaginal access is needed for natural orifice transluminal endoscopic surgery. We introduce a new culdotomy procedure, using a technique for the creation of space in the cul-de-sac, transvaginal ultrasound, and a newly developed umbrella Hakko needle. An artificially developed, saline-containing space in the cul-de-sac was punctured by an umbrella Hakko needle from the vagina under the guidance of ultrasound. The vaginal walls on both sides of the needle were incised with an electric scalpel. In five cases with benign gynecological tumors, culdotomy was successfully performed. Operating time was less than 10 minutes and blood loss was less than 10 mL. There were no culdotomy-associated complications. This procedure, named Culdotomy FourS Two U, is a simple, safe, and reliable method for entry into the cul-de-sac in transvaginal gynecological surgeries and may have future applications in transluminal endoscopic surgery through the vagina.

Ultrasound-guided culdotomy for vaginal ovarian cystectomy using a renal balloon dilator catheter.

OBJECTIVES: To evaluate the feasibility and utility of performing ultrasound-guided culdotomy using a renal balloon dilator catheter for transvaginal ovarian cystectomies. METHODS: Culdotomy using transvaginal sonography and a Nephromax balloon dilator catheter was performed in 16 patients for the vaginal removal of benign ovarian cysts located in the cul-de-sac. Each ovarian cyst was punctured under transvaginal ultrasound guidance and the punctured site on the vaginal wall was enlarged with a dilator. The cyst was then enucleated through this vaginal wound. Preoperative characteristics of the patients, outcome, operating time, blood loss and complications of each culdotomy, and the histology of the cysts, were recorded and examined. RESULTS: We used this method on 14 patients with unilateral ovarian cysts and two with bilateral cysts. Culdotomy was performed successfully in 15 cases (94%). The mean +/- SD operating time for culdotomy was 22 +/- 11 min, and blood loss during the procedure was less than 10 mL in all cases. There were no complications including rectal injury or febrile morbidity. CONCLUSIONS: Culdotomy assisted by ultrasound imaging and a dilator is a safe, reliable and effective method for removal of benign ovarian cysts via a vaginal approach.

Lymphokine-independent, leukemia cell-mediated induction of tumor necrosis factor in human monocytes.

Unactivated human blood monocytes and monocytic THP-1 cells were found to respond to some leukemia cells by tumor necrosis factor (TNF) production. The TNF production by THP-1 cells in response to K562 cells was preceded by a rapid rise in [Ca2+]i, initiated within 1 h and terminated within 4 h as a refractory state took over. Neither the amount nor the duration of TNF production was enhanced by gamma-interferon. The P32/ISH cells did not induce a significant [Ca2+]i change of TNF production, while MOLT-4 cells failed to induce TNF despite their capacity to mobilize Ca2+ in THP-1 cells. The failure of P32/ISH or MOLT-4 to induce TNF was attributed primarily to a lack of stimulatory membrane molecules rather than to suppression by an inhibitory component, since liposomes carrying membrane components of K562 and MOLT-4 or P32/ISH in varying proportions elicited TNF production that precisely reflected the K562 proportion. The ability of K562 to induce TNF was selectively impaired by trypsin, whereas the ability to mobilize [Ca2+]i was more sensitive to glutaraldehyde, although once the latter activity was extinguished, the K562 cell could no longer induce TNF. These results suggest that some leukemia cells are equipped with two or more signaling membrane moieties which together stimulate monocytes for transient tumoricidal expression in the preimmune stage.

Novel mutation (E113X) of antithrombin III gene (AT3) in a woman with gestational recurrent thrombosis.

A 35-year-old Japanese woman with a low level (42-54%) of blood antithrombin (AT) III, experienced two induced abortions due to deep venous thrombosis at 8 weeks of gestation (GW) and cerebral thrombosis at 10 GW. The present pregnancy was successfully managed with intravenous administration of AT III (6,000-8,000 U/wk). Analysis of polymerase chain reaction (PCR)-single strand conformation polymorphism (SSCP) for exons 3A and 4 of the AT III gene (AT3) using her DNA revealed extra expansion bands with altered migration. The DNA sequencing demonstrated novel mutations in exon 3A of AT3: a G to T substitution at nucleotide position 5333 in codon GAG for Glu 113, causing a stop codon (E113X), and an A to T substitution at position 5338 in codon AAA for Lys 114, forming Asn (K114N). These novel mutations, especially E113X, in AT3 may be related to recurrent thrombosis in the pregnancy.

Recent trends in the prevalence of Down syndrome in Japan, 1980-1997.

The aims of the present study were to determine recent trends in the prevalence of Down syndrome (DS) in Japan, and to determine whether recent changes in demographic and social habits and access to prenatal diagnosis have influenced the livebirth rates of DS. Livebirth statistics indicate that the birth rate in Japan has decreased for women in their 20s and has increased for those in their 30s and 40s. During an 18-year period between 1980 and 1997, 1,299 consecutive DS infants were born among a total of 2,232,694 births, a rate corresponding to approximately 10% of all births in Japan over the same period. The increasing risk of DS with advancing maternal age was confirmed. The overall prevalence was 5.82 DS births per 10,000 livebirths (8.3-9.7 per 10,000 after correction according to the estimated ascertainment ratio: 60-70%). The prevalence rate by year of child birth represents a statistically significant increase (P = 0.001). In conclusion, recent trends in the prevalence of DS in Japan from 1980 to 1997 failed to show a consistent tendency to decrease, probably because of the concomitant increase in pregnancy in advanced maternal age.

Use of PCR serum in diagnosing and monitoring cytomegalovirus reactivation in bone marrow transplant recipients.

We previously reported that the use of polymerase chain reaction (PCR) in detecting cytomegalovirus (CMV) DNA in serum (sPCR) enables the detection of CMV viremia, which has not been possible with other methods. In this study, the clinical usefulness of sPCR was investigated by comparison with the results of three other diagnostic methods, i.e., antigenemia assay (AG), shell vial culture test (shell vial), and complement-fixing (CF) antibody titer. The present study included 26 patients with hematological diseases who had undergone allogeneic bone marrow transplantation (BMT). A total of 347 samples were collected, and the results of the sPCR and AG methods were in agreement in 91.1% of the samples. When a subject was positive in both the sPCR and AG tests, and the other two tests (shell vial and CF) were also positive, CMV reactivation was surmised as definite. When only the result of the shell vial test or the CF test was positive, these results were taken as false-positives. The time at which the samples became positive in each of these four tests was 7.5 weeks post-BMT for sPCR, 7.0 weeks post-BMT for the AG test, 7.4 weeks post-BMT for the shell vial test, and 9.7 weeks post-BMT for the CF test. Thus, it was found that samples became positive at almost the same time for the sPCR, AG, and shell vial tests. Interstitial pneumonitis (IP) due to CMV developed in 3 subjects. These cases were positive in the sPCR, AG, and shell vial tests prior to the manifestation of symptoms of IP. The CF test did not become positive until after the onset of the disease. As the IP due to CMV was controlled with treatment, the sPCR and AG tests became negative. With the shell vial and CF tests, on the other hand, the test results continued to be positive even after the IP was cured. These findings demonstrate that the sPCR test method--like the AG test--yields few false-positive results. Therefore, the sPCR method is useful in early diagnosis of reactivation of CMV and for evaluation of the efficacy of therapy administered for IP. In addition, sPCR can be performed simultaneously on a large number of samples, and the evaluation of the test results is simple. We conclude that the sPCR test may be superior to the three other diagnostic methods for evaluation of serum samples from multiple institutions.

Tubal pregnancy in a unicornuate uterus with rudimentary horn: a case report.

OBJECTIVE: To report a case of tubal pregnancy in a unicornuate uterus with rudimentary horn on the side of the rudiment and its pathology. DESIGN: Case report. SETTING: University hospital. PATIENT: An 18-year-old woman, primigravida, with tubal pregnancy. INTERVENTION(S): Systemic administration of methotrexate, salpingectomy by laparotomy, and laparoscopic surgery for resection of rudimentary horn. MAIN OUTCOME MEASURE(S): Emergent laparotomy revealed that the intraperitoneal hemorrhage was caused by the rupture of the tubal pregnancy on the same side as the rudimentary horn of the unicornuate uterus. A corpus luteum was found at the ipsilateral ovary of the rudimentary horn. RESULT(S): Image diagnosis and pathological examination of the rudimentary horn revealed that this uterine malformation was a unicornuate uterus with a noncommunicated, noncavitary rudimentary horn, corresponding to class IIc of the American Fertility Society classification of müllerian anomalies. CONCLUSION(S): This is the first report of a tubal pregnancy on the side of the noncommunicating rudimentary horn with the ipsilateral ovary carrying a corpus luteum in a unicornuate uterus.

Enzyme-linked immunosorbent assay (ELISA) for human epidermal growth factor (hEGF).

An enzyme-linked immunosorbent assay for human epidermal growth factor (hEGF) by the sandwich method with the use of orthophenylene diamine (OPDA) for the substrate of the enzyme reaction was developed. This assay showed high sensitivity, and there was no problem with the recovery rates even without any pretreatment of samples. The changes of hEGF levels in plasma and serum depend on the condition of blood storage from sampling until separation of the cell content. The levels of hEGF markedly increased with time in serum, especially when stored at room temperature but less so in plasma. Changes of hEGF levels in plasma were negligible for up to 3 h if the blood was stored at 4 degrees C until separation.

Alpha-fetoprotein microheterogeneity: a potential biochemical marker for Down's syndrome.

Our purpose was to examine the utility of analyzing alpha-fetoprotein (AFP) microheterogeneity assessed by lectin affinity in Down's syndrome (DS) screening. Maternal sera and amniotic fluids were collected from 18 women who were carrying DS fetuses and 70 unaffected pregnancies around 16 weeks of gestation. The percentages of AFP which reacted with Lens culinaris agglutinin (AFP-L2,3) were determined by lectin affinity electrophoresis. AFP-L2,3 levels were significantly increased (P<0.0001) in both maternal serum and amniotic fluid from DS-affected versus unaffected pregnancies. The fractional areas under the receiver operating characteristic curves were 0.835 and 0.700 (P=0.106) for AFP-L3 and AFP MoM (multiples of the median) in maternal serum. No correlation was found between AFP-L3 and AFP MoM in maternal serum (r=0.006). Our data suggest that the measurement of AFP-L3 in maternal serum is a potential biochemical marker for DS.

Retention versatility of silica-supported comb-shaped crystalline and non-crystalline phases in high-performance liquid chromatography.

Silica-supported comb-shaped polymer (Sil-ODA18) provided a specific selectivity towards aromatic and non-aromatic solutes. Selectivity of Sil-ODA18 and ODS (polymeric and monomeric) columns towards polycyclic aromatic hydrocarbons and geometric isomers have been discussed in respect of molecular shapes and sizes. The retention versatility of this phase arises from the orientation change on transition from the ordered crystalline to the less ordered non-crystalline state. Selectivity in the crystalline and non-crystalline regions of Sil-ODA18 is similar to that of polymeric and monomeric ODS phases, respectively. Sil-ODA18 column also exhibited the characteristic temperature dependency of liquid-crystalline phases, i.e., an increase in retention with increasing temperature on transition from crystalline to isotropic state.

Intraperitoneal administration of cytomegalovirus hyperimmunoglobulin to the cytomegalovirus-infected fetus.

Twenty-five percent of cytomegalovirus (CMV)-infected fetuses had sequelae and 8% of those in the recurrent-infected group had sequelae. There is no report yet on the fetal therapy for CMV infections. A Japanese pregnant woman with intrauterine fetal CMV infection diagnosed at 26 weeks of pregnancy is presented. CMV culture of amniotic fluid was positive. A CMV DNA assay using the polymerase chain reaction method of the cord blood and the amniotic fluid was positive during the pregnancy; however, testing for fetal serum CMV-specific IgM was negative. The CMV IgG titer of fetal serum at 27 weeks of pregnancy was a third of that of the maternal serum. CMV hyperimmunoglobulin was injected into the fetal abdominal cavity at 28 and 29 weeks of pregnancy. A second administration of CMV hyperimmunoglobulin increased the titer of CMV IgG in the fetal circulation. At birth, the urine culture was positive for CMV. However, CMV DNA of the ascites became negative. A brain CT scan performed 2 weeks after birth revealed some small calcifications beside the right ventricle. CMV hyperimmunoglobulin injection to the fetal abdominal cavity has been shown to increase the IgG in the fetal serum. This is the first report of fetal therapy of congenital CMV infection.

Spontaneous reseal of the fetal membranes in patients with high-leak PROM, confirmed by intra-amniotic injection of a dye (phenol-sulfonphthalein).

Two cases (27 and 23 weeks of gestation) diagnosed as high-leak PROM by an intra-amniotic dye (PSP) injection method later proved negative on re-testing by the same method, and ended in spontaneous reseal of the membranes. The pregnancies in these cases extended to 36 and 40 weeks of gestation without any eventful signs of infection.

Diagnosis of preterm premature rupture of the membranes using a newly developed AFP monoclonal antibody test kit.

OBJECTIVE: We have developed a new anti-AFP monoclonal antibody kit which is easier to use than other examination methods for detecting AFP in the leaked amniotic fluid. In this study, we investigate the clinical value of this test in the diagnosis of preterm premature rupture of the membranes (PROM). METHODS: We employed 103 patients of less than 37 weeks of gestational age for this preliminary study. We compared the clinical usefulness of this new AFP test with that of the nitrazine test and measured the concentration of AFP in vaginal fluid or cervical secretion by EIA. RESULTS: The nitrazine test showed a correct diagnostic rate of 62.1%, in contrast the AFP test kit had a 98.0% rate (P < 0.001). The reaction time using the kit is 3 min. CONCLUSION: The AFP test is a simple and non-invasive test which can be easily carried out repeatedly as a bedside examination. This study has confirmed the high clinical efficacy of the newly developed AFP test kit as a method of PROM diagnosis.

Diagnosis of premature rupture of the membranes in preterm patients, using an improved AFP kit: comparison with ROM-check and/or nitrazine test.

OBJECTIVE: We developed a new kit for detecting AFP in leaked amniotic fluid. Later, we developed an improved AFP kit utilizing the same anti alpha-fetoprotein (AFP) monoclonal antibody. In this study, we evaluate the clinical usefulness of this improved kit in the diagnosis of preterm premature rupture of membranes (PROM). METHODS: We compared this improved AFP test with the ROM-check and/or the nitrazine tests in 46 preterm patients. RESULTS: The ROM-check and nitrazine tests showed a diagnostic accuracy of 89.1 and 87.0%, respectively, compared with 95.7% with the improved AFP test. The sensitivity of the improved AFP test on cervical samples was significantly higher than that of the nitrazine test on vaginal samples (P < 0.05). The reaction time with the improved AFP kit test is 90 s. CONCLUSION: This study has confirmed a great clinical utility of the improved AFP test kit as a method of PROM diagnosis.

A case of intrauterine medical treatment for cystic hygroma.

We report the first case of an intrauterine treatment for cystic hygroma. Guided by ultrasonography, we first removed intracystic fluid from two cysts and then injected OK-432 into each fetal cyst at 21 and 28 weeks of gestation. No re-enlargement of the cysts was subsequently observed. At 38 weeks of gestation, a male infant was delivered transvaginally. Only a slight skin fold was observed in the nuchal area of the neonate, indicating the effectiveness of OK-432 for the intrauterine treatment of cystic hygroma.