Feasibility of transperineal minimal invasive surgery when performing sacrectomy for advanced primary and recurrent pelvic malignancies.

BACKGROUND: This study aimed to clarify the efficacy and safety of minimally invasive transabdominal surgery (MIS) with transperineal minimal invasive surgery (tpMIS) for sacrectomy in advanced primary and recurrent pelvic malignancies. METHODS: Using a prospectively collected database, we retrospectively analyzed the clinical, surgical, and pathological outcomes of MIS with tpMIS for sacrectomies. Surgery was performed between February 2019 and May 2023. The median follow-up period was 27 months (5-46 months). RESULTS: Fifteen consecutive patients were included in this analysis. The diagnoses were as follows: recurrent rectal cancer, n = 11 (73%); primary rectal cancer, n = 3 (20%); and recurrent ovarian cancer, n = 1 (7%). Seven patients (47%) underwent pelvic exenteration with sacrectomy, six patients (40%) underwent abdominoperineal resection (APR) with sacrectomy, and two patients (13%) underwent tumor resection with sacrectomy. The median intraoperative blood loss was 235 ml (range 45-1320 ml). The postoperative complications (Clavien-Dindo grade ≥ 3a) were graded as follows: 3a, n = 6 (40%); 3b, n = 1 (7%); and ≥ 4, n = 0 (0%). Pathological examinations demonstrated that R0 was achieved in 13 patients (87%). During the follow-up period, two patients (13%) developed local re-recurrence due to recurrent cancer. The remaining 13 patients (87%) had no local disease. Fourteen patients (93%) survived. CONCLUSIONS: Although the patient cohort in this study is heterogeneous, MIS with tpMIS was associated with a very small amount of blood loss, a low incidence of severe postoperative complications, and an acceptable R0 resection rate. Further studies are needed to clarify the long-term oncological feasibility.

Feasibility of transanal minimally invasive surgery for total pelvic exenteration for advanced primary and recurrent pelvic malignancies.

BACKGROUND: The purpose of this study was to clarify the efficacy and safety of transanal minimally invasive surgery (TAMIS) for total pelvic exenteration (TPE) in advanced primary and recurrent pelvic malignancies. METHODS: Using a prospectively collected database, we retrospectively analyzed the clinical, surgical, and pathological outcomes of TAMIS for TPE. Surgery was performed between September 2019 and April 2023. The median follow-up period was 22 months (2-45 months). RESULTS: Fifteen consecutive patients were included in this analysis M:F = 14:1 and median (range) age was 63 (36-74). Their diagnoses were as follows: primary rectal cancer (n = 5; 33%), recurrent rectal cancer (n = 4; 27%), primary anorectal cancer (n = 5; 33%), and gastrointestinal stromal tumor (n = 1; 7%). Bladder-sparing TPE was selected for two patients (13%). In nine of 15 patients (60%) the anal sphincter could be successfully preserved, five patients (33%) required combined resection of the internal iliac vessels, and two (13%) required rectus muscle flap reconstruction. The median operative time was 723 min (561-1082), and the median intraoperative blood loss was 195 ml (30-1520). The Clavien-Dindo classifications of the postoperative complications were as follows: grade 0-2 (n = 11; 73%); 3a (n = 3; 20%); 3b (n = 1; 7%); and ≥ 4 (n = 0; 0%). No cases of conversion to laparotomy or mortality were observed. The pathological results demonstrated that R0 was achieved in 14 patients (93%). CONCLUSIONS: The short-term outcomes of this initial experience proved that this novel approach is feasible for TPE, with low blood loss, acceptable postoperative complications, and a satisfactory R0 resection rate.

DISC1 regulates cell-cell adhesion, cell-matrix adhesion and neurite outgrowth.

Disrupted-in-schizophrenia 1 (DISC1) is a promising susceptibility gene for major mental illness. Recent studies have implicated DISC1 in key neurodevelopmental processes, including neurite outgrowth, neuronal migration and proliferation. Here, we report that DISC1 regulates cell-cell and cell-matrix adhesion and neurite outgrowth. DISC1 overexpression increased expression of the adherence junction protein N-cadherin and enhanced cell-cell adhesion. The increased N-cadherin accumulated in the areas of cell-cell contact. DISC1 overexpression also enhanced cell-matrix adhesion by inducing expression of beta1-integrin protein. In the presence of nerve growth factor (NGF), DISC1 overexpression increased beta1-integrin expression at the cell membrane and growth cone. NGF-induced neurite extension was enhanced by DISC1, and anti-beta1-integrin antibody reduced the neurite outgrowth of DISC1-overexpressing cells to the control level. Furthermore, DISC1 also regulated N-cadherin and beta1-integrin expression at the cell membrane in primary neurons. We conclude that DISC1 regulates cell-cell adhesion and cell-matrix adhesion by regulating the expression of adhesion molecules.

Clear evidence for element partitioning effects in a Ti-6Al-4V alloy by the first-principles phase field method.

Ti-6 wt% Al-4 wt% V (Ti64) is an α + ß titanium alloy, in which the alloying components strongly affect the mechanical properties. In this report, element partitioning effects in Ti64 are investigated by using the first-principles phase field (FPPF) method, which has recently been proposed by our group. In the FPPF method, the local free energy is calculated using a cluster expansion method in combination with density functional theory and the temperature effect is incorporated using potential renormalization theory. We have succeeded in identifying enrichment of Al (V) in the α (ß) phase, i.e., the clear evidence for the element partitioning effects of Al and V, without using any thermodynamical parameter. The transformation of the ß phase and the α phase in microstructure is investigated by varying the V and Al concentrations by a small amount. Our results are in excellent agreement with the recent experimental results, showing the validity of the FPPF method for ternary alloys.

Neurotransmitter release from synaptotagmin-deficient clonal variants of PC12 cells.

Synaptotagmin (p65) is an abundant synaptic vesicle protein of neurons and contains regions similar to the regulatory domain of protein kinase C. These domains are thought to be involved in calcium-dependent interaction with membrane phospholipids during exocytosis. To assess the functional role of synaptotagmin, synaptotagmin-deficient clonal variants of PC12 cells were isolated. All of the variant cells released catecholamine and adenosine triphosphate in response to elevated intracellular concentrations of calcium, which suggests that synaptotagmin is not essential for secretion of catecholamine and adenosine triphosphate from PC12 cells.

Establishment of variant PC12 subclones deficient in stimulation-secretion coupling.

Clonal rat pheochromocytoma (PC12) cells have been widely used to study the molecular mechanism of exocytosis. We have isolated variant PC12 subclones with deficiencies in stimulation-secretion coupling, by a single cell recloning, and investigated the defects. PC12-1G2 hardly released dopamine following high-K(+)-induced depolarization, but normal release was evoked by the Ca(2+)-ionophore, ionomycin. Fura-2 fluorometry indicated that a nicardipine-sensitive component of Ca(2+) influx was missing, suggesting that PC12-1G2 has defects in L-type Ca(2+) channel function. PC12-2B3 was not responsive to high-K(+)-induced depolarization and ionomycin, and voltage-dependent Ca(2+) entry was identical to that of the normal clone. Electron microscopy revealed that the number of vesicles adjacent or directly attached to the plasma membrane was decreased in PC12-2B3. The expression of presynaptic proteins was analyzed by immunoblotting using a panel of antibodies. Syntaxin 1, VAMP-2, SNAP-25, Munc18, Rab3C and Sec-6 were decreased compared to the control clone and that of synaptophysin was extremely low. PC12-D60 synthesized and released dopamine normally, but had almost lost its catecholamine-uptake activity. These results show that multiple PC12 cells variants are spontaneously generated, and that recloning can select PC12 subclones useful for the study of the molecular mechanisms of neurotransmitter release.

Latent tuberculous infection in the United States and Canada: who completes treatment and why?

OBJECTIVES: To assess latent tuberculous infection (LTBI) treatment completion rates in a large prospective US/Canada multisite cohort and identify associated risk factors. METHODS: This prospective cohort study assessed factors associated with LTBI treatment completion through interviews with persons who initiated treatment at 12 sites. Interviews were conducted at treatment initiation and completion/cessation. Participants received usual care according to each clinic's procedure. Multivariable models were constructed based on stepwise assessment of potential predictors and interactions. RESULTS: Of 1515 participants initiating LTBI treatment, 1323 had information available on treatment completion; 617 (46.6%) completed treatment. Baseline predictors of completion included male sex, foreign birth, not thinking it would be a problem to take anti-tuberculosis medication, and having health insurance. Participants in stable housing who received monthly appointment reminders were more likely to complete treatment than those without stable housing or without monthly reminders. End-of-treatment predictors of non-completion included severe symptoms and the inconvenience of clinic/pharmacy schedules, barriers to care and changes of residence. Common reasons for treatment non-completion were patient concerns about tolerability/toxicity, appointment conflicts, low prioritization of TB, and forgetfulness. CONCLUSIONS: Less than half of treatment initiators completed treatment in our multisite study. Addressing tangible issues such as not having health insurance, toxicity concerns, and clinic accessibility could help to improve treatment completion rates.

Synaptotagmin I is essential for Ca(2+)-independent release of neurotransmitter induced by alpha-latrotoxin.

alpha-Latrotoxin causes massive release of norepinephrine from clonal rat pheochromocytoma PC12 cells, in the absence of external Ca2+, by an unknown mechanism. The effect almost disappeared in PC12 variant cells deficient in synaptotagmin I, a synaptic vesicle protein, and was rescued by transfecting the synaptotagmin I gene. These results indicate that synaptotagmin I is essential for the Ca(2+)-independent action of alpha-latrotoxin in PC12 cells.

Regulation of nerve growth factor and nerve growth factor receptor production by NMDA in C6 glioma cells.

The synthesis of nerve growth factor (NGF) and nerve growth factor receptor (NGFR) were studied in a C6 glioma cell line by Northern blot hybridization. In response to a glutamate agonist N-methyl-D-aspartic acid (NMDA), NGF mRNA increased by up to 2-fold after 4-12 h of culture. The non-NMDA receptor agonists, quisqualate and kainate, did not induce any increase of NGF mRNA, and kainate actually produced a decrease. The increase in NGF mRNA in response to NMDA was dose-dependent at 1, 5 and 10 microM. NGF receptor (NGFR) mRNA showed changes in expression which were similar to those for NGF mRNA, but were less marked. The specific glutamate antagonist 2-aminophosphonovaleric acid (APV) blocked the increase of NGF mRNA produced by NMDA. In the absence of Ca2+, an increase of NGF mRNA was still observed but in the presence of 1 mM ethylglycol-bis-(beta-aminoethyl ether) N,N'-tetraacetic acid (EGTA), NGF mRNA production abolished. The mechanism producing an increase in NGF mRNA by NMDA may be mediated by cyclic AMP since intracellular cyclic AMP and NGF mRNA levels both increased following treatment with NMDA or dibutyryl cyclic AMP.

Effect of unusual polyamines on cleavage of DNA by restriction enzymes.

The effect of unusual polyamines, such as thermine, caldopentamine, caldohexamine, tris(3-aminopropyl)amine, or tetrakis(3-aminopropyl)ammonium, on the activities of various restriction endonucleases was investigated by using an Escherichia coli plasmid as a substrate, which contains a high GC content fragment from an extreme thermophile. Restriction enzymes used were SmaI, BanII, NaeI, RsaI, and TaqI. Most of the polyamines tested were inhibitory to the enzyme activities. The larger and more branched a polyamine was, the more the activities of nucleases were inhibited. The inhibition was positively correlated with the polyamine concentration. The sites protected by a polyamine were identical to those protected by other polyamines, and also identical to those which were less sensitive to the restriction enzyme in the absence of polyamines. No sequence specificity was seen among these sites.

Factors associated with identifying tuberculosis contacts.

SETTING: Little is known about why some tuberculosis (TB) patients identify few or even no contacts. OBJECTIVES: To describe patient perceptions of the contact investigation interview and determine potential factors associated with identifying TB contacts. DESIGN: A total of 13 focus groups were conducted: 10 groups with previously smear-positive pulmonary TB patients born in the United States or Mexico, and three with program staff to discuss attitudes toward and perceptions of the contact investigation interview. Patients were recruited into separate groups by country of birth and number of contacts identified. RESULTS: The data indicated that patients-even those who identified few contacts-overwhelmingly reported identifying contacts easily and willingly. Understanding the purpose of the contact investigation and seriousness of TB facilitated naming contacts, while miscommunication and misconceptions about TB hindered the process. Patients felt strongly about informing their contacts before the health department contacted them. Staff respondents reported that education and effective communication were critical during contact investigation interviewing. CONCLUSION: Data indicated that patients, including those identifying few contacts, reported wanting to name their contacts. However, misconceptions may affect their understanding of who their contacts are, and hence the quantity and quality of the contacts identified. These findings underscore the need for effective communication and education.

Synthesis of kanamycin A analogs containing 6-amino-3-oxa-2,3,4,6-tetradeoxy-D- and -L-glycero-hexopyranose.

6-Azido-3-oxa-2,3,4,6-tetradeoxy-D- and -L-glycero-hexopyranoses were synthesized in five steps from (2S)-and (2R)-1,2-O-isopropylideneglycerols, respectively. After conversion into the corresponding ethyl 1-thioglycosides, each was condensed with a protected derivative of 6-O-(3-amino-3-deoxy-alpha-D-glucopyranosyl)-2-deoxystreptamine (16). Deprotection and reduction of the azido group of the condensation products gave the title compounds.

Synthesis of dibekacin analogs containing 3-oxa- and 3-aza-2,3,4-trideoxy-D-glycero-hexopyranose.

6-O-(3-Oxa-2,3,4-trideoxy-alpha-D-glycero-hexopyranosyl) derivatives (10 and 17) of both 3',4'-dideoxyneamine and 5-epifluoro-5,3',4'-trideoxyneamine have been prepared by coupling ethyl 6-O-benzyl-3-oxa-2,3,4-trideoxy-1-thio-D-glycero-hexopyranoside (5) with suitable aglycons. The corresponding 3"-aza derivative (19) of dibekacin (6) was prepared by oxidation of 1,3,2',6'-tetra-N-tosyldibekacin (7) with Pb(OAc)4 followed by treatment with NH4OAc and reduction with NaBH3CN. Related ring-opening compounds (11 and 25) were also prepared.

TB perspectives among a sample of Mexicans in the United States: results from an ethnographic study.

OBJECTIVE: A study was conducted to describe the sociocultural aspects of tuberculosis (TB) among Mexicans in the U.S. and to provide TB programs with practical recommendations for serving this population. METHODS: In-depth, structured, open-ended interviews were conducted with 50 persons from Mexico living in the U.S. Local bilingual, bicultural researchers conducted the interviews with respondents recruited from TB clinics and surrounding communities. Both qualitative and quantitative strategies were used to analyze the data. RESULTS: We found diverse TB perceptions and attitudes, but few were associated with specific participant characteristics. We detected widespread misperceptions about TB transmission and low perceptions of risk. Anticipated TB stigma among those with no history of disease was qualitatively greater than reported stigma among those who had TB disease. We also detected missed opportunities for TB education. Reported barriers to care included lack of transportation, limited clinic hours, cost of services, inconvenient clinic location, and communication problems with staff. CONCLUSIONS: To address the diverse needs of Mexican-born clients, we recommend that TB programs provide culturally-appropriate, patient-centered care. We suggest several strategies aimed at raising risk awareness and reducing stigma. Finally, we encourage programs to facilitate access by providing language-appropriate services, extending clinic hours, and facilitating transportation.

Transfer of spermatogenesis-related cDNAs into eel testis germ-somatic cell coculture pellets by electroporation: methods for analysis of gene function.

Genes encoding spermatogenesis-related substance (eSRSs) show unique expression patterns during spermatogenesis. To analyze their function, we developed a new assay system using gene transfer techniques combined with coculture of the eel germ-somatic cells. First, we investigated the efficacy of in vitro electroporation transfer of gene into germ-somatic cell pellets using green fluorescent protein (GFP) gene. Second, in order to define the function of the eSRSs, we electrophoretically transferred eel spermatogonial stem cell renewal factor (eSRS34) and eel spermatogenesis-preventing substance (eSRS21) genes into germ-somatic cell pellets. Presence of the transferred cDNA was examined by reverse transcription-polymerase chain reaction (RT-PCR). Furthermore, proliferating cells were detected histologically, after labeling with BrdU. Transfer of the eSRS34 gene induced spermatogonial stem cell renewal in the pellets. Moreover, 11-ketotestosterone (11-KT) treatment stimulated the proliferation of spermatogonia, which resulted in the appearance of late type B spermatogonia in the pellets. The proliferation of spermatogonia by 11-KT stimulation was suppressed by transfer of the eSRS21 gene. These results indicate that the transferred eSRS34 and 21genes were functional in the pellets. Thus, an efficient in vitro gene transfer technique for coculture system of germ and somatic cell of Japanese eel was established.

Emla versus ice as a topical anesthetic.

BACKGROUND: There are many different topical anesthetics available. Ice is an easy to use and inexpensive topical anesthetic. There are studies comparing different topical anesthetics but none have used ice. OBJECTIVE: To evaluate the efficacy between EMLA (2.5% lidocaine and 2.5% prilocaine cream) and ice prior to injection with a 30-gauge needle containing a painful stimuli. METHOD: A fingertip amount of EMLA cream was placed on the upper left arm and occluded with a bandage for an average of 52 minutes. Ice in a rubber glove was placed on the right upper arm for 1-2 minutes. The EMLA site, ice site, and a control site without anesthetic were tested for cutaneous analgesia with an injection of 1% lidocaine with 1:100,000 epinephrine through a 30-gauge needle using a 3 cc syringe. Discomfort was recorded using a visual acuity scale (VAS). Analysis of data was performed using a t test paired sample of two means. RESULTS: Statistically there was a significant difference in pain control between EMLA and ice (P <.05), EMLA and control (P <.01), and ice and control (P <.01). Clinically patients felt that EMLA performed slightly better as an analgesic than ice, and both EMLA and ice were more efficacious than the control. CONCLUSION: Both EMLA and ice decreased the discomfort associated with needle injection. Although EMLA performed better in pain control, ice has advantages in ease of use, fast action, and is less expensive than EMLA. Both EMLA and ice were good topical anesthetics, each with advantages and disadvantages in clinical use.

Forceps and cotton applicator method of freezing benign lesions.

BACKGROUND: Cryosurgery is one of the most common treatment methods in the dermatologists armamentarium. We describe a method combining forceps and a cotton applicator to freeze benign lesions for practitioners who prefer to use cotton-tipped applicators instead of a cryosurgery spray device. METHODS: We describe the technique using common in-office instruments. CONCLUSION: Lesions around delicate areas can be treated with forceps and a cotton-tipped applicator dipped in liquid nitrogen resulting in less pain and decreased collateral damage vs. traditional methods. For the most delicate areas, the forceps dipped directly in liquid nitrogen method is preferred.

Cyclic adenylic acid-dependent and -independent production of chloramphenicol acetyltransferase in Escherichia coli carrying R plasmids.

A plasmid-specified, inducible, but cyclic adenylic acid (cAMP)-independent resistance to chloramphenicol (CM) is reported. The resistance level to CM was increased two- to fourfold by treatment with a small amount of CM for a short period in Escherichia coli strains carrying the R plasmid pJY1, which was obtained from a clinical isolate of Vibrio cholerae. Though cAMP was required for production of CM acetyltransferase (CATase) in cAMP-deficient mutants of E. coli carrying the R100 plasmid, the same species harboring pJY1 did not require cAMP for production of the enzyme. The possibility of a mechanism other than CATase activity for the CM resistance conferred by R plasmids is discussed.