Modified open posterior internal sphincterotomy with sliding skin graft for chronic anal fissure and anal stenosis: Low recurrence rate and no serious faecal incontinence postoperative complication.

AIM: Lateral internal sphincterotomy (LIS) remains a standard for chronic anal fissure even though other surgical techniques have shown high efficacy. Faecal incontinence is a well-documented complication of LIS. We devised modified open posterior internal sphincterotomy (m-OPIS) with sliding skin graft (SSG), which is a combined procedure of OPIS and anal advancement flap. The aim of this study is to evaluate m-OPIS+SSG. METHODS: This was a retrospective, observational, single-arm study. m-OPIS+SSG was performed for chronic anal fissure and anal stenosis. m-OPIS involved incision of the internal sphincter muscle at the posterior midline until four fingers could be passed. The incision wound was closed by anastomosis of the anoderm and skin. Then, an arcuate skin incision was created and the skin graft was advanced into the anal canal. Follow-up was conducted by clinical consultation and telephone interview. Faecal continence was assessed by Cleveland Clinic Faecal Incontinence (CCFI) score. RESULTS: m-OPIS+SSG was performed in 143 patients. The mean patient age was 50±16 years. The success and overall recurrence rates after m-OPIS+SSG were 99% and 0.7%, respectively, with a median follow-up period of 16.3 years. One patient developed incontinence with liquid stools once during the 6-month period. None of the other patients suffered permanent faecal incontinence postoperatively. The postoperative CCFI score was 0.5±0.9. CONCLUSIONS: We consider m-OPIS+SSG as one of the efficacious options of procedure for chronic anal fissure and anal stenosis, owing to its high success rate, low recurrence rate and no postoperative complication of serious faecal incontinence.

Usefulness and complications associated with thenar and standard portals during arthroscopic surgery of thumb carpometacarpal joint.

PURPOSE: Advances in small arthroscopy have enabled a minimally invasive surgery for thumb carpometacarpal joints. However, surgery is often difficult using standard CM-radial (CM-R) and CM-ulnar portals (CM-U). Here, we describe the clinical applications and complications associated with using thenar portal (TP) and standard portals. METHODS: Arthroscopic surgeries of thumb carpometacarpal joint were performed in 21 patients including 15 patients with osteoarthritis and six Bennett's fracture-dislocations. Complications and the frequency of use associated with each portal were evaluated. RESULTS: Complications associated with the CM-R portal comprised paresthesia due to damage of the radial nerve branches in two patients. No nerves were damaged but the operation scar became tender at the TP in three patients. The CM-R was used at a lower frequency when the TP was utilized. CONCLUSION: The clinical use of TP may decrease the risk of radial sensory nerve damage through decreasing frequency of use of the CM-R that is located near the nerve. LEVEL OF STUDY: IV.

High lifetime and reproductive performance of sows on southern European Union commercial farms can be predicted by high numbers of pigs born alive in parity one.

Our objectives were 1) to compare reproductive performance across parity and lifetime performance in sow groups categorized by the number of pigs born alive (PBA) in parity 1 and 2) to examine the factors associated with more PBA in parity 1. We analyzed 476,816 parity records and 109,373 lifetime records of sows entered into 125 herds from 2008 to 2010. Sows were categorized into 4 groups based on the 10th, 50th, and 90th percentiles of PBA in parity 1 as follows: 7 pigs or fewer, 8 to 11 pigs, 12 to 14 pigs, and 15 pigs or more. Generalized linear models were applied to the data. For reproductive performance across parity, sows that had 15 or more PBA in parity 1 had 0.5 to 1.8 more PBA in any subsequent parity than the other 3 PBA groups ( P< 0.05). In addition, they had 2.8 to 5.4% higher farrowing rates in parities 1 through 3 than sows that had 7 or fewer PBA (P < 0.05). However, there were no differences between the sow PBA groups for weaning-to-first-mating interval in any parity (P ≥ 0.37). For lifetime performance, sows that had 15 or more PBA in parity 1 had 4.4 to 26.1 more lifetime PBA than sows that had 14 or fewer PBA (P < 0.05). Also, for sows that had 14 or fewer PBA in parity 1, those that were first mated at 229 d old (25th percentile) or earlier had 2.9 to 3.3 more lifetime PBA than those first mated at 278 d old (75th percentile) or later (P < 0.05). Factors associated with fewer PBA in parity 1 were summer mating and lower age of gilts at first mating (AFM; P < 0.05) but not reservice occurrences (P = 0.34). Additionally, there was a 2-way interaction between mated month groups and AFM for PBA in parity 1 (P < 0.05); PBA in parity 1 sows mated from July to December increased nonlinearly by 0.3 to 0.4 pigs when AFM increased from 200 to 310 d old (P < 0.05). However, the same rise in AFM had no significant effect on the PBA of sows mated between January and June (P ≥ 0.17). In conclusion, high PBA in parity 1 can be used to predict that a sow will have high reproductive performance and lifetime performance. Also, the data indicate that the upper limit of AFM for mating between July and December should be 278 d old.

Interactions between pre- or postservice climatic factors, parity, and weaning-to-first-mating interval for total number of pigs born of female pigs serviced during hot and humid or cold seasons.

The objective of this study was to examine interactions between climatic factors, parity, and weaning-to-first-mating interval (WMI) for total number of pigs born at subsequent parity (TPB) of female pigs serviced during 2 seasons. The present study analyzed records of 27,739 gilts and 127,670 parity records of sows in 95 Japanese herds; the records included females that were serviced between June and September (hot and humid season) or between December and March (cold season) in 2007 through 2009. The climate data were obtained from 20 weather stations located close to the studied herds. Mean daily maximum temperatures (Tmax), mean daily minimum temperatures (Tmin), and daily average relative humidity (RH) for 21 d preservice and 15 d postservice for each female were coordinated with that female's reproductive data. Linear regression models with random intercept and slopes were applied to the data. Mean TPB (±SEM) was 11.9 ± 0.01 pigs. Mean values (ranges) of Tmax in the hot and humid season and Tmin in the cold season were 28.4 (13.6 to 39.8°C) and 2.0°C (-13.2 to 17.6°C), respectively. Also, mean RH in the hot and humid season and the cold season were 73.2 (35 to 98%) and 65.2% (25 to 99%), respectively. In the hot and humid season, TPB in gilts decreased by 0.05 pigs for each degree Celsius increase in preservice Tmax (P < 0.05). However, there was no association between gilt TPB and either postservice Tmax (P = 0.11) or pre- and postservice RH (P ≥ 0.66). In sows, as preservice Tmax increased from 25 to 30°C, TPB in parity groups 1 and 2 or higher decreased by 0.6 and 0.4 pigs, respectively (P < 0.05). Also, sow TPB decreased by 0.1 to 0.4 pigs as postservice Tmax increased from 25 to 30°C (P < 0.05). In sows with WMI of 0 to 12 d, TPB decreased by 0.2 to 0.5 pigs as pre- or postservice Tmax increased from 25 to 30°C (P < 0.05). However, in sows with WMI of 13 d or more, TPB was not associated with pre- or postservice Tmax (P ≥ 0.10). As preservice Tmax increased from 25 to 30°C, TPB in sows under 81.6% RH (90th percentile) decreased by 0.5 pigs (P < 0.05), whereas TPB in sows under 65.7% RH (10th percentile) decreased by only 0.3 pigs (P < 0.05). Postservice RH in the hot and humid season was not associated with sow TPB (P = 0.18). During the cold season there was no association between TPB and pre- or postservice Tmin (P ≥ 0.09) or RH (P ≥ 0.45). Therefore, we recommend that producers apply cooling management for females during periservice in summer to increase TPB.

Bispectral index is related to the spread of spinal sensory block in patients with combined spinal and general anaesthesia.

BACKGROUND: A relationship between the depth of sedation as measured by the bispectral index (BIS) and spinal sensory block height in patients with light to no additional sedation has been described previously. The present study was designed to investigate the hypothesis that BIS values closely correlate with the spread of spinal sensory block in patients deeply sedated with an i.v. target-controlled infusion of propofol. METHODS: Subjects comprised 100 patients aged 20-64 yr and undergoing arthroscopic knee surgery. Patients were given spinal anaesthesia with bupivacaine 0.5% (3 ml). Propofol was administered to achieve a target effect-site concentration of 3.0 µg ml⁻¹. The relationship between the spinal sensory level at 15 min after spinal anaesthesia and BIS values during 1-5, 6-10, 11-15, and 16-20 min time intervals after the estimated effect-site concentration reached 3.0 µg ml⁻¹ was evaluated. RESULTS: The sensory level of spinal analgesia significantly and strongly correlated with BIS values during each time period after the estimated effect-site concentration remained at 3.0 µg ml⁻¹ (P<0.0001). The correlation coefficient values were 0.8 during 1-5 min, 0.844 during 6-10 min, 0.801 during 11-15 min, and 0.804 during 16-20 min time periods. CONCLUSIONS: We demonstrated that BIS values significantly correlate with the level of spinal sensory block under deep sedation with propofol. The depth of sedation induced by spinal anaesthesia depends on the spread of spinal sensory block.

A single amino acid substitution can shift the optimum pH of DNase I for enzyme activity: biochemical and molecular analysis of the piscine DNase I family.

We purified four piscine deoxyribonucleases I (DNases I) from Anguilla japonica, Pagrus major, Cryprus carpio and Oreochromis mossambica. The purified enzymes had an optimum pH for activity of approximately 8.0, significantly higher than those of mammalian enzymes. cDNAs encoding the first three of these piscine DNases I were cloned, and the sequence of the Takifugu rubripes enzyme was obtained from a database search. Nucleotide sequence analyses revealed relatively greater structural variations among the piscine DNase I family than among the other vertebrate DNase I families. From comparison of their catalytic properties, the vertebrate DNases I could be classified into two groups: a low-pH group, such as the mammalian enzymes, with a pH optimum of 6.5-7.0, and a high-pH group, such as the reptile, amphibian and piscine enzymes, with a pH optimum of approximately 8.0. The His residue at position 44 of the former group is replaced by Asp in the latter. Replacement of Asp44 of piscine and amphibian DNases I by His decreased their optimum pH to a value similar to that of the low-pH group. Therefore, Asp44His might be involved in an evolutionarily critical change in the optimum pH for the activity of vertebrate DNases I.

Five age-dependently expressed genes in mouse brain revealed by the fluorescence differential display-PCR technique.

We used a fluorescence differential display-PCR (FDD-PCR) technique to analyze the genes expressed in mouse brains collected at nine different developmental stages ranging from 3 days to 15 months after birth, and 5 age-dependently expressed genes were found. Age-dependent expression of each of these 5 genes was confirmed by quantitative real-time PCR analysis. Of the 5 genes, 4 (B1-B4) had high homology with the nucleotide sequences of cDNA clones of known mouse genes (myelin proteolipid protein, transferrin, embryo cDNA from the RIKEN full-length enriched library, and protein tyrosine phosphatase), and the rest (B5) with expressed sequence tags of an unknown gene. Sequencing analysis of the full-length cDNA constructed based on the B5 sequence demonstrated that the gene product of B5 was identical to G-substrate, a specific substrate for cGMP-dependent protein kinase. The expression patterns of known genes obtained in our study may provide a further opportunity to investigate the biological and physiological roles of the proteins they encode.

Characterization and haplotype analysis of the polymorphic Y-STRs DYS443, DYS444 and DYS445 in a Japanese population.

From sequence database information we have newly identified three male-specific and polymorphic tetranucleotide STRs, DYS443 (GDB: 10807127), DYS444 (GDB: 10807128) and DYS445 (GDB: 10807129) on the Y chromosome. Analysis of 190 Japanese males revealed 6, 5 and 4 alleles in the DYS443, DYS444 and DYS445 systems, with calculated STR diversities of 0.68, 0.57 and 0.53, respectively. The cumulative haplotype diversity of the five Y-STRs DYS441, DYS442, DYS443, DYS444 and DYS445 was calculated to be 0.95 and therefore application of these STRs may yield very useful information for forensic individualization.

Identification and characterization of two novel human polymorphic STRs on the Y chromosome.

From sequence database information, we have identified two male-specific and polymorphic tetranucleotide STRs, DYS 441 (GDB:10013873) and DYS 442 (GDB: 10030304), on the Y chromosome. Analysis of 184 males allowed 7 and 5 alleles to be distinguished in the DYS 441 and DYS 442 systems, respectively, yielding 21 haplotypes. The gene diversities were 0.72 and 0.51, respectively and the haplotype diversity was 0.85.

Amphibian DNases I are characterized by a C-terminal end with a unique, cysteine-rich stretch and by the insertion of a serine residue into the Ca2+-binding site.

We purified four amphibian deoxyribonucleases I from the pancreases of one toad, two frog and one newt species, by using three different column chromatography methods in sequence. Each of the purified enzymes had a molecular mass of approx. 40 kDa and an optimal pH for activity of approx. 8.0. These values were significantly greater than those for other vertebrate DNases I. The full-length cDNA encoding each amphibian DNase I was constructed from the total RNA of the pancreas by using rapid amplification of cDNA ends. Nucleotide sequence analyses revealed two structural characteristics unique to amphibian DNases I: a stretch of approx. 70 amino acids with a high cysteine content (approx. 15%) in the C-terminal region, and the insertion of a serine residue at position 205 (in a domain containing an essential Ca2+-binding site). Expression analysis of a series of mutant constructs indicated that both of these structures are essential in generating the active form of the enzyme. 'DNase I signature sequences', which are well conserved in other vertebrate DNases I, could not be found in any of the amphibian DNases I tested, whereas a 'somatomedin B motif' was identified in the Cys-rich stretches of all four. Although DNase I has so far been considered to be a secretory glycoprotein, amphibian DNase I seems to be non-glycosylated. These structural findings indicate strongly that amphibian DNases I are situated in a unique position on the phylogenetic tree of the DNase I family.

Cloning, mapping, genomic organization, and expression of mouse M-LP, a new member of the peroxisomal membrane protein Mpv17 domain family.

We have identified a mouse full-length cDNA and gene encoding a novel protein (M-LP), based on an expressed sequence tag (EST) sequence (GenBank Accession No. AI482564) obtained by differential display screening of age-dependently expressed genes in mouse kidney. The ML-P gene is composed of three exons, ranges over 5 kb on mouse chromosome 16B1-B2 and is expressed as two transcripts (1455 and 3058 bp), both of which include the same open-reading frame encoding 194 amino acids. M-LP is expressed mainly in kidney and spleen and shows age-dependent expression. M-LP has sequence homologies and membrane topologies very similar to the Mpv17 protein, a peroxisomal protein involved in the development of early-onset glomerulosclerosis. Search of the protein domain family database (ProDom) revealed that M-LP is a new member of the Mpv17 domain family (PD008400).

Tissue-specific in vivo inhibition of DNase I gene expression by somatostatin.

Administration of somatostatin to rats induced a transient reduction of serum levels of deoxyribonuclease I (DNase I) activity in a dose-dependent manner, followed by a substantial decrease of DNase I activity in the lower gut. Activity in the parotid gland, liver, and kidney did not change. Real-time PCR analysis of the DNase I gene transcript in ileum indicated that the decrease was due to down-regulation of gene expression. Based on these responses, rat tissues expressing DNase I could be classified into two types, somatostatin-sensitive and somatostatin-resistant, and the level of DNase I activity in the lower gut seems to be controlled by somatostatin.

Use of human recombinant DNase I expressed in COS-7 cells as an immunogen to produce a specific anti-DNase I antibody.

To obtain human deoxyribonuclease I (DNase I) as an immunogen, we have developed a procedure that is more useful and effective than the conventional procedure, which uses human urine as a starting material. In the new procedure, we culture COS-7 cells transfected with expression vector carrying human DNase I cDNA, and then purify the enzyme from the culture medium. The enzyme can be easily isolated to apparent homogeneity by passage through only three chromatography columns. The rabbit antiserum that we used against the recombinant DNase I was not inferior to that used against DNase I from human urine, in terms of both its ability to discriminate DNase I phenotypes and its ability to neutralize enzyme activity. Therefore, our procedure may be useful for producing an antibody specific for human DNase I.

Neurobehavioral alterations in mice with a targeted deletion of the tumor necrosis factor-alpha gene: implications for emotional behavior.

Tumor necrosis factor-alpha (TNF-alpha) is emerging as an important modulator of the function of the central nervous system. In the present study, we investigated a role of endogenous TNF-alpha in cognitive and emotional function using mice with targeted deletions of the TNF-alpha gene. TNF-alpha-(-/-) mice showed normal diurnal rhythms of spontaneous locomotor activity and cognitive functions. Emotional behavior in the mutant mice, however, was significantly altered, which manifested in the performance in the open-field, elevated plus maze, and forced swimming tests. The altered performance in the elevated plus maze test was significantly alleviated by treatment with diazepam. Postmortem brain analysis of TNF-alpha-(-/-) mice revealed a significant increase in serotonin metabolism in the brain. These findings suggest a role for endogenous TNF-alpha in emotional behavior, which may possibly be related to alterations of serotonine metabolism.

Suppression of neurocognitive damage in LP-BM5-infected mice with a targeted deletion of the TNF-alpha gene.

Brain levels of TNF-alpha increase in many inflammatory conditions, including HIV-1 infection, and may contribute to neurodegenerative processes. The paucity of agents that can selectively and potently block TNF-alpha processing or its receptors has led us to investigate the role of TNF-alpha in chronic neurodegeneration associated with retroviral infection using mice with targeted deletions of the TNF-alpha gene. Infection of wild-type C57BL/6 mice with the LP-BM5 murine leukemia retrovirus mixture leads to the development of a severe immunodeficiency as well as cognitive deficits and neuronal damage. TNF-alpha-(-/-) mice infected with LP-BM5 developed a systemic immunopathology indistinguishable in severity from that observed in contemporaneously infected wild-type mice. In contrast, the performance of infected TNF-alpha-(-/-) mice in the Y-maze and Morris water maze was not different from that of uninfected TNF-alpha-(-/-) mice. The extent of glial activation in the striatum, as indicated by the increase in density of peripheral benzodiazepine receptors, was equivalent in both groups of LP-BM5-infected mice. However, the decrease in striatal MAP-2 expression, a marker of neurodegeneration observed in infected wild-type mice, was not found in infected TNF-alpha-(-/-) mice. While the loss of TNF-alpha appeared to have no effect on the course or severity of the central or peripheral immunopathology resulting from LP-BM5 infection, the behavioral and biochemical manifestations were substantially curtailed in the TNF-alpha-(-/-) mice. These findings directly support a role for TNF-alpha in the neurodegenerative processes associated with viral infections such as HIV-1.

Age-related changes of gene expression in mouse kidney: fluorescence differential display--PCR analyses.

We used a fluorescence differential display--PCR (FDD-PCR) technique to analyze the genes expressed in mouse kidneys collected at nine different developmental stages ranging from 3 days to 15 months after birth. We found ten genes that were age-dependent and differentially-expressed in the kidneys during our experimental period. We confirmed by comparative RT-PCR that of the ten cDNAs, seven showed reproducible age-dependent expression. Four of the nucleotide sequences of these cDNA clones, had high homology with known genes (fibronectin, soluble guanylyl cyclase alpha-1 subunit, cytosolic aldehyde dehydrogenase and mitochondrial DNA), and three with expressed sequence tags of unknown genes. The FDD-PCR method was very useful for detecting new age-related genes expressed differentially in the mouse kidney.

Japanese population data for two STR loci, HumTPO and HumLPL.

The two short tandem repeat (STR) systems, HumTPO and HumLPL, were investigated in blood samples obtained from approximately 800 unrelated Japanese individuals living in seven geographically different areas of Japan. Neither deviation from a Hardy-Weinberg equilibrium nor significant difference between the allele distributions was found among the seven Japanese populations in the two STR systems. These findings indicate that there is a general uniformity for both the STR loci in the Japanese population.