Single Ligation Research Articles

Synergistic protective effects of erythropoietin and olmesartan on ischemic stroke survival and post-stroke memory dysfunctions in the gerbil

Treatment with erythropoietin and AT1 blockers is protective in experimental acute cerebral ischemia, with promising results in pilot clinical studies in human stroke. This paper examines the effects of using both agents as combination therapy in acute ischemic stroke. We used the single carotid ligation stroke model in the gerbil. Six groups of 50 gerbils were treated either with placebo, erythropoietin (intraperitoneally, 5000 IU/kg, 2 and 48 h after stroke), olmesartan (10 mg/kg per day in drinking water started 36 h after stroke), ramipril (2.5 mg/kg per day in drinking water started 36 h after stroke), erythropoietin + olmesartan, or erythropoietin + ramipril. Long-term (1 month) Kaplan-Meyer survival curves were obtained, and survivors were submitted at day 30 to immediate (object recognition test) and spatial (Morris water maze) memory function tests. Erythropoietin alone and olmesartan alone, but not ramipril, significantly increased survival at day 30 compared with untreated controls (38, 30 and 6% versus 12%, respectively). Combined treatment with erythropoietin and olmesartan further increased the survival rate to 56%, whereas combined therapy with erythropoietin and ramipril decreased 30-day survival to 24% (P < 0.0001, erythropoietin + olmesartan versus erythropoietin + ramipril). Untreated stroke survivors had markedly altered performances in both the object recognition test (P = 0.0007) and the Morris water maze (P < 0.0001) tests at day 30 compared with normal gerbils. In erythropoietin-treated animals, ramipril therapy had no beneficial effect whereas olmesartan fully restored normal response to the memory tests. Post-infarct treatment with olmesartan combined with early erythropoietin therapy has a protective effect on survival, and markedly improves long-term memory dysfunction in this experimental model.

Complexation of Copper(II)−Chelidamate: A Multifrequency-Pulsed Electron Paramagnetic Resonance and Electron Nuclear Double Resonance Analysis

Multifrequency electron paramagnetic resonance (EPR) and electron nuclear double resonance (ENDOR) techniques were used to obtain structural information about the copper(II)-chelidamate complex. Well-resolved nitrogen ENDOR spectra could be recorded from solid solution samples by using selective excitation of spin packets. Evaluation of nuclear quadrupole and dipolar hyperfine interaction of the directly ligated nitrogen allowed for an identification of the bond direction to the copper ion within the eigen frame of the copper g-matrix. Invoking two-dimensional EPR techniques, additional hyperfine interaction with a "distant" nitrogen spin, identified as resulting from the solvent dimethylformamide (DMF), was observed. The experimental data are only consistent with formation of a stable pseudoplanar copper complex with single solvent ligation via its oxygen atom.

A simplified method for cloning of short interfering RNAs from Brassica juncea infected with Turnip mosaic potyvirus and Turnip crinkle carmovirus

RNA silencing is a plant defense mechanism in which virus infected plants produce short interfering RNAs (siRNAs) derived from viral RNA, that attack the virus at the post-transcriptional level. In a previous study on Cymbidium ringspot tombusvirus (CymRSV) infection in Nicotiana benthamiana, siRNAs (determined by cloning and sequencing) predominantly originated from the sense (+) strand of the viral RNA, suggesting that the majority of siRNAs are produced through the direct cleavage of the virus single strand (ss) RNA by the plant Dicer-like enzyme. To test whether this asymmetry in strand polarity is a generic rule for all plant viruses, siRNAs from Brassica juncea, either singly infected by Turnip mosaic potyvirus (TuMV, the family Potyviridae), or doubly infected with TuMV and Turnip crinkle carmovirus (TCV, the family Tombusviridae) were investigated. A simplified siRNA cloning method was developed, using a single ligation reaction to attach both 5′ and 3′ adapters to the target short RNAs followed by one-step RT-PCR amplification. In the TCV infection, as for the CymRSV infection, siRNAs were produced predominantly (97.6%) from the +ss RNA. However, for TuMV infections, siRNAs were derived from both strands (+/−, 58.1–41.9%), indicating the presence of alternative siRNA production mechanisms.

A genomic library-based amplification approach (GL-PCR) for the mapping of multiple IS 6110 insertion sites and strain differentiation of Mycobacterium tuberculosis

Evidence suggests that insertion of the IS 6110 element is not without consequence to the biology of Mycobacterium tuberculosis complex strains. Thus, mapping of multiple IS 6110 insertion sites in the genome of biomedically relevant clinical isolates would result in a better understanding of the role of this mobile element, particularly with regard to transmission, adaptability and virulence. In the present paper, we describe a versatile strategy, referred to as GL-PCR, that amplifies IS 6110-flanking sequences based on the construction of a genomic library. M. tuberculosis chromosomal DNA is fully digested with HincII and then ligated into a plasmid vector between T7 and T3 promoter sequences. The ligation reaction product is transformed into Escherichia coli and selective PCR amplification targeting both 5′ and 3′ IS 6110-flanking sequences are performed on the plasmid library DNA. For this purpose, four separate PCR reactions are performed, each combining an outward primer specific for one IS 6110 end with either T7 or T3 primer. Determination of the nucleotide sequence of the PCR products generated from a single ligation reaction allowed mapping of 21 out of the 24 IS 6110 copies of two 12 banded M. tuberculosis strains, yielding an overall sensitivity of 87,5%. Furthermore, by simply comparing the migration pattern of GL-PCR-generated products, the strategy proved to be as valuable as IS 6110 RFLP for molecular typing of M. tuberculosis complex strains. Importantly, GL-PCR was able to discriminate between strains differing by a single IS 6110 band.

Infected femoral pseudoaneurysm in intravenous drug abusers.

The result of surgical treatment of 21 infected femoral pseudoaneurysms in 19 intravenous drug addicts was evaluated. Eight pseudoaneurysms involved only the common or superficial femoral artery and 13 involved the femoral bifurcation. Excision and ligation was performed as the sole procedure in 19 instances, and revascularization by bypass through the obturator route was carried out in two. The mean follow-up was 12.3 months. One patient required an above-knee amputation. The resultant ischaemia was greater after triple vessel ligation (mean ankle:brachial pressure index (ABPI) 0.41) than single vessel ligation (mean ABPI 0.58). Postoperative bleeding occurred in one patient. Intermittent claudication was present in 14 patients after excision and ligation. Claudication was universal and more severe after triple than after single vessel ligation. There was no subsequent limb loss. Excision and ligation is safe and is the treatment of choice for infected femoral pseudoaneurysm in drug addicts.

A New Model of Peripheral Arterial Disease: Sustained Impairment of Nutritive Microcirculation and Its Recovery by Chronic Electrical Stimulation

To develop a model of peripheral arterial disease (PAD) in rat skeletal muscle with sustained impairment of microcirculatory perfusion, and to ascertain whether increased muscle activity can reverse the impairment. Three weeks after iliac ligation in rats, the ipsilateral femoral artery was ligated (double ligation, DL), and in some animals, muscle activity was increased by electrical stimulation for 2 weeks (10 Hz, 15 min on, 85 mins off, 7 times per day). Diameter changes of precapillary arterioles to vasoactive agonists and capillary perfusion (flow intermittency, capillary red cell velocity [V(rbc)], and diameters) were measured in extensor digitorum longus muscle and compared with 5 weeks iliac only ligation (single ligation, SL) and controls. Total muscle endothelial nitric oxide synthase (eNOS) was estimated by Western blotting. Whereas single ligation increased intermittency of capillary flow with little effect on V(rbc) and shear stress, DL completely eliminated increases in V(rbc) and shear stress after muscle contractions. Arterial dilation to sodium nitroprusside was attenuated similarly in SL and DL; in SL, acetylcholine induced constriction and bradykinin an attenuated dilation, but in DL vessels were unresponsive to either. Chronic stimulation returned all microcirculatory parameters in DL to normal and increased levels of eNOS protein by 75%. Femoral artery ligation following iliac ligation impairs arteriolar vasodilator capacity, capillary perfusion, and shear-dependent function of microcirculatory endothelium more than iliac ligation alone and is more representative of long-standing ischemia in PAD. Chronic intermittent electrical stimulation can normalize these derangements.

Endoscopic ligation for bleeding esophageal varices: Comparison of single ligation and spiral multi-ligation

Endoscopic ligation for bleeding esophageal varices: Comparison of single ligation and spiral multi-ligation

Chronic Portal Hypertension in the Rat by Triple-Portal Stenosing Ligation

A surgical technique based on the development of a triple stenosing ligation is used to worsen the complications inherent to the prehepatic chronic portal hypertension. The results have been compared with those obtained in rats with a single-portal stenosing ligation. An increase ( p < .05) in the body, liver, spleen, and kidney weights as well as a decrease ( p < .001) in the testes weight to body weight ratio were produced in both groups of animals. In addition, the variability in the obtained weights, particularly in the liver weight, stands out. The incidence of portosystemic and portohepatic collateral circulation and of the mesenteric venous vasculopathy increases in the animals with triple-portal stenosing ligation. The new proposed technique is a valid alternative to the classic one that used single portal stenosing ligation.

The influence of DNA double-strand break structure on end-joining in human cells.

DNA end-joining is the major repair pathway for double-strand breaks (DSBs) in higher eukaryotes. To understand how DSB structure affects the end-joining process in human cells, we have examined the in vivo repair of linearized plasmids containing complementary as well as several different configurations of non-complementary DNA ends. Our results demonstrate that, while complementary and blunt termini display comparable levels of error-free rejoining, end-joining fidelity is decreased to varying extents among mismatched non-complementary ends. End structure also influences the kinetics of repair, accurately recircularized substrates for blunt and complementary termini being detected significantly earlier than for mismatched non-complementary ends. These results suggest that the end-joining process is composed of an early component, capable of efficiently repairing substrates requiring a single ligation event, and a late component, involved in the rejoining of complex substrates requiring multiple processing steps. Finally, these two types of repair events may have different genetic requirements as suggested by the finding that exposure of cells to wortmannin, a potent inhibitor of phosphatidylinositol 3-related kinases (PI 3-related kinases), blocks the repair of complex substrates while having little or no effect on those requiring a simple ligation event.

Coordination chemistry of ( c-C 5H 5)Fe + in the gas phase at 294 ± 3 K: reactions with the inorganic ligands H 2, H 2O, NH 3, CO, N 2, NO, CO 2, N 2O, and NO 2

Experimental results are reported for primary and higher-order gas-phase reactions of ( c-C 5H 5)Fe + with H 2, H 2O, NH 3, CO, N 2, NO, CO 2, N 2O, and NO 2. Reaction rate coefficients and product distributions were measured with the selected-ion flow tube technique operating at 294 ± 3 K and a helium buffer-gas pressure of 0.35 ± 0.01 Torr. The measurements provide intrinsic efficiencies for ligation and lead to intrinsic coordination numbers based on ligation kinetics. The extent of sequential ligation was found to be strongly dependent on the nature of the ligand. Single ligation of the ( c-C 5H 5)Fe + cation was observed with H 2, N 2, N 2O, and CO 2 whereas multiple ligation was observed with NO, NO 2, CO, H 2O, and NH 3. With NO 2 oxygen-atom transfer was the dominant reaction channel. Structures have been proposed for the ligated ions and relative bond energies were investigated using multicollision induced dissociation.

Increased Duodenal Mucosa Infiltration by Mast Cells in Rats with Portal Hypertension

Background: Enteropathy characterized by vascular and inflammatory alterations in the submucosa and mucosa has been described in patients with portal hypertension. Aims: To verify the theory of inflammatory etiopathogenesis in experimental portal hypertensive duodenopathy, a prehepatic portal hypertension model based on the development of a single and triple partial ligation of the portal vein was used in the rat. Methods: Five rats in each group (male Wistar, 230–255 g) were subjected to single (group II) or triple (group III) partial ligation of the portal vein and then compared to 5 control animals (group I, no operation). The animals were sacrificed 6 weeks later to analyze the histological parameters of the duodenal mucosa and submucosa, i.e., number, diameter and area of submucosal vessels, density of mast cells and mitotic cells. Body, liver and spleen weights and collateral circulation type were also assayed. Results: As was demonstrated by the collateral circulation in all of the animals, the partial portal ligation was successful. Compared to the controls, the number of vessels per microscopic field (25 ± 3.16 vs. 18.60 ± 1.52), their diameter (20.09 ± 2.90 vs. 12.61 ± 3.97 µm, p < 0.05) and consequently their total area (12,749.30 ± 2,298.26 vs. 3,455.82 ± 1,702.33 µm²) were increased in the animals with a single partial ligation (group II) as well as in animals receiving triple partial ligation (group III) (33 ± 12.88, p < 0.05; 22.92 ± 6.72 µm, p < 0.05 and 51,376.95 ± 43,732.24 µm², p < 0.05, respectively). In addition, the density of mast cells increased from 3.26 ± 1.18 in controls to 10.74 ± 1.47, p < 0.01 and 22.50 ± 6.42, p < 0.01 in single and triple partial portal ligated animals, respectively. Mitosis was significantly induced in crypts of the duodenal mucosa of the single portal ligated animals (25.20 ± 1.78 vs. 17.40 ± 1.14, p < 0.01) but was inhibited in triple partial ligated animals (12.40 ± 5.12, p < 0.05). Compared to controls, both groups of rats developed liver atrophy with a greater decrease in the liver/body weight ratio in the single (2.71 ± 0.50%, p < 0.01) compared to the triple partial ligated animals (3.33 ± 0.09%, p < 0.01). Conclusions: The correlation of the degree of portal hypertension with the vascular changes and mast cell density suggests that both the hypertensive state and inflammation may play a role in the development of portal hypertensive intestinal vasculopathy. The inverse relation of portal hypertension with liver atrophy and mitosis rate in the crypts of the duodenal mucosa has not been clarified and should be investigated in future studies.

Long-term results of rubber-band ligation for second-degree haemorrhoids: a prospective study

Rubber-band ligation is a widely performed method of treatment for second-degree haemorrhoids. Initial results of rubber-band ligation are satisfactory, but symptoms often recur in the long term. The aim of our prospective study was to present the long-term results of rubber-ring application in patients with second-degree haemorrhoids. From 1990 to 1996, 114 outpatients with second-degree haemorrhoids underwent rubber-band ligation according to the Barron technique. A single ligation was performed during one session. Most of the patients had three haemorrhoids being ligated. Three years after the initial treatment, 105 patients were re-examined and their symptoms were recorded. 16 patients underwent an additional rubber-band ligation 6–24 months after their initial treatment. After 3 years, 30 patients (28.5%) were symptom-free, 17 patients (16.2%) were improved, 32 patients (30.5%) were unchanged and 26 patients (24.8%) were worse. Bleeding and prolapse were the most frequent recurrent symptoms. Rubber-band ligation is a simple, safe and inexpensive method of treatment for second-degree haemorrhoids, although symptoms may recur in many patients in the future.

Gas-Phase Coordination of Mg+, (c-C5H5)Mg+, and (c-C5H5)2Mg+ with Saturated Hydrocarbons

The coordination of the electronic ground states of Mg+, (c-C5H5)Mg+, and (c-C5H5)2Mg+ with the straight-chain saturated hydrocarbons methane, ethane, n-propane, n-butane, n-pentane, n-hexane, and n-heptane has been investigated in the gas phase in helium at room temperature and moderate pressures. Reaction rate coefficients and product distributions were measured with the selected-ion flow tube (SIFT) technique operating at 294 ± 3 K and a helium buffer-gas pressure of 0.35 ± 0.01 Torr. Rate coefficients were measured for all observed ligation steps (or upper limits in the case of nonreactions), and bond connectivities in the coordinated ions were probed with multicollision-induced dissociation. Only single ligation was observed. The rate of ligation was found to depend on the size of the saturated hydrocarbon, increasing with increasing size until the collision rate is reached, and on the degree of ligation with c-C5H5. Mg+ was found to be unreactive with methane and ethane, k < 10-13 cm3 molecule-1 s-1, but ligation was observed with the other hydrocarbons, k ≥ 7 × 10-12 cm3 molecule-1 s-1. Single ligation of Mg+ with c-C5H5 substantially enhances the efficiency of subsequent ligation: the ligation is rapid, k ≥ 1.4 × 10-10 cm3 molecule-1 s-1, with all the saturated hydrocarbons investigated. Double ligation with c-C5H5 completely shuts down the efficiency of ligation: no reaction was observed between the saturated hydrocarbons and the full-sandwich magnesocene cation, k < 10-13 cm3 molecule-1 s-1. A linear correlation is reported between the measured thresholds for multicollision-induced dissociation and the polarizability of the hydrocarbon ligand. With a view to published calculations by Bauschlicher et al. for Mg+−CH4 and Mg+−C2H4, this correlation is consistent with end-on structures for the Mg+−L and (c-C5H5)Mg+−L species observed in this study. A detailed study using density functional theory for the interaction of Mg+ with three rotamers of n-pentane indicates that interconversion of different isomers of Mg+−n-C5H12 should be easy, even at room temperature and that the ligated species Mg+−n-C5H12 is “dynamic” rather than “static”. Stronger bonding (by a factor of about two) is indicated for (c-C5H5)Mg+−L by the multicollision CID experiments, and this is consistent with a formal description of (c-C5H5)Mg+ as (c-C5H5)-Mg2+.

A binary vector-based large insert library forBrassica napusand identification of clones linked to a fertility restorer locus forOguracytoplasmic male sterility (CMS)

We constructed and characterized a large DNA insert library for Brassica napus that would facilitate genome-related research and map-based cloning efforts in Brassica species. This library, consisting of 92 160 clones arrayed in 384-well microtiter dishes, was based on a conventional plant transformation vector (binary vector), and was constructed using a single ligation with transformation efficiency of over 5000 recombinants per microliter of ligation mixture. Every clone in this library contains an insert in the size range of 30-190 kb, facilitating both chromosome walking and plant transformation. Screening this library with three DNA markers (C2, F10, and CabR) that are linked to a fertility restorer locus for Ogura cytoplasmic male sterility (CMS) identified at least 17 positive clones for each probe. Among the 17 positive clones identified by C2, nine are linked to the restorer locus. Marker F10 identified 21 clones, of which only two are linked to the restorer locus. None of 68 clones identified by CabR is linked to the restorer locus. A stability test using two clones identified by the C2 marker indicated that large DNA inserts are stable in this conventional vector in both Escherichia coli and Agrobacterium. Key words: Brassica napus, binary vector, large DNA insert library, restorer gene.

Rubber Band Ligation of Symptomatic Internal Hemorrhoids: Results of 500 Cases

Background/Aim: In this prospective study the results of rubber band ligation (RBL) of symptomatic hemorrhoids in 500 consecutive patients with 2nd (255 cases), 3rd (218 cases) and 4th degree (27 cases) hemorrhoids are presented. Methods: The patients’ symptoms were hemorrhage in 142 cases (28.4%), prolapse in 33 cases (6.6%) and both hemorrhage and prolapse in 325 cases (65%). Sixteen patients with hemorrhoids had liver cirrhosis and portal hypertension. RBL was performed using the St Marks’ applicator (Seward) on an outpatient basis. Multiple ligations in two (259 cases) or three (190 cases) sessions were undertaken in 449 patients (89.8%), while a single ligation was done in 51 cases (10.2%). Results: Successful results were achieved in 440 cases (88%) in a 24-month follow-up. A total of 94 patients (18.8%) had complications which required no hospitalization. Pain and hemorrhage were the most frequent complications. RBL proved to be safe in 16 patients with coagulation disorders due to liver cirrhosis. Two years after RBL, symptomatic recurrence was 11.9% (53/445) with repeat RBL or surgery in 9.2% (41/445). Conclusions: RBL is a useful, safe and successful method for treating symptomatic 2nd and 3rd degree hemorrhoids, which can be applied successfully in selected cases with 4th degree hemorrhoids, but with an increased rate of recurrence and additional treatment requirements. Also, RBL seems to be safe in patients with liver cirrhosis and portal hypertension.

A binary vector-based large insert library for &lt;i&gt;Brassica napus&lt;/i&gt; and identification of clones linked to a fertility restorer locus for &lt;i&gt;Ogura&lt;/i&gt; cytoplasmic male sterility (CMS)

We constructed and characterized a large DNA insert library for Brassica napus that would facilitate genome-related research and map-based cloning efforts in Brassica species. This library, consisting of 92,160 clones arrayed in 384-well microtiter dishes, was based on a conventional plant transformation vector (binary vector), and was constructed using a single ligation with transformation efficiency of over 5000 recombinants per microliter of ligation mixture. Every clone in this library contains an insert in the size range of 30-190 kb, facilitating both chromosome walking and plant transformation. Screening this library with three DNA markers (C2, F10, and CabR) that are linked to a fertility restorer locus for Ogura cytoplasmic male sterility (CMS) identified at least 17 positive clones for each probe. Among the 17 positive clones identified by C2, nine are linked to the restorer locus. Marker F10 identified 21 clones, of which only two are linked to the restorer locus. None of 68 clones identified by CabR is linked to the restorer locus. A stability test using two clones identified by the C2 marker indicated that large DNA inserts are stable in this conventional vector in both Escherichia coli and Agrobacterium.

Gas-Phase Coordination of Mg+, (c-C5H5)Mg+, and (c-C5H5)2Mg+ with Small Inorganic Ligands

The coordination of the electronic ground states of Mg+, (c-C5H5)Mg+, and (c-C5H5)2Mg+ with the inorganic molecules H2, NH3, H2O, N2, CO, NO, O2, CO2, N2O, and NO2 has been investigated in the gas phase at room temperature and moderate pressures. Reaction rate coefficients and product distributions were measured with the selected-ion flow tube (SIFT) technique operating at 294 ± 3 K and a helium buffer-gas pressure of 0.35 ± 0.01 Torr. Rate coefficients were measured for all observed ligation steps (or upper limits in the case of nonreactions), and bond connectivities in the coordinated ions were probed with multicollision-induced dissociation. The rates and extent of ligation were found to depend on the degree of ligation with c-C5H5 and the nature and size of the inorganic ligand. Mg+ was found to be unreactive except with ammonia, which was found to add slowly (k = 4 × 10-12 cm3 molecule-1 s-1). Single ligation of Mg+ with c-C5H5 substantially enhances the efficiency of ligation; the initial ligation is rapid (k > 5 × 10-11 cm3 molecule-1 s-1) with all ligands except H2, N2, and O2. Double ligation with c-C5H5 substantially reduces the efficiency of ligation; no ligation was observed with the “full-sandwich” magnesocene cation. Fast bimolecular ligand-switching reactions occur instead, but only with NH3 and H2O, indicating that these two molecules ligate more strongly to (c-C5H5)Mg+ than does c-C5H5 itself. Higher-order ligation was observed between Mg+ and NH3 and between the “half-sandwich” (c-C5H5)Mg+ cation and CO, NO, NH3, H2O, CO2, N2O, and NO2. After a slow initiation step, ammonia added to Mg+ up to four times to generate Mg(NH3)4+. Higher-order ligation reactions of (c-C5H5)Mg+ with CO, NO, NH3, CO2, N2O, and NO2 proceed with diminishing rates adding up to at most three ligands. H2O is an exception; at least six H2O molecules were observed to add sequentially to (c-C5H5)Mg+. Variations in the observed rate of this addition have been attributed to the completion of an “inner” coordination shell and the onset of hydrogen bonding in an “outer” coordination shell that accounts for the continuing rapid ligation. Higher-order ligands were removed sequentially by multicollision-induced dissociation with He atoms; there was no evidence for intramolecular interligand interactions leading to unimolecular bond redisposition after ligation.

Construction and Characterization of a Bacterial Artificial Chromosome Library of the Indica Rice Kasalath.

We constructed a bacterial artificial chromosome (BAC) library for the indica rice variety Kasalath using a new BAC vector, pCHR8, suited for plasmid rescue, containing nine unique restriction sites in a multicloning site. This Kasalath BAC Iibrary consists of about 65, 000 clones with an average insert size of 106 kb using HindIII partially digested DNA fragments. It represents approximately sixteen rice haploid genome equivalents which are sufficient for the full coverage of the rice genome with BAC clone contigs. The library was constructed from only a single ligation mixture with high transformation efficiency. The quality of this BAC Iibrary was ascertained by successfully retrieving the BAC clones for seven rice RFLP markers. We could isolate eleven individual BAC clones using RFLP markers that linked to the lethal factor locus for segregation distortion on rice chromosome 3. The content of the chloroplast and mitochondrial DNA in this library was estimated to be 2.8% and 0.4%, respectively. These results demonstrate that this Kasalath BAC Iibrary should be suitable for map-based cloning and physical mapping of the rice genome.

Intrahepatic STAT-3 activation and acute phase gene expression predict outcome after CLP sepsis in the rat.

Interleukin-6 (IL-6) regulates hepatic acute phase responses by activating the transcription factor signal transducer and activator of transcription (STAT)-3. IL-6 also may modulate septic pathophysiology. We hypothesize that 1) STAT-3 activation and transcription of alpha2-macroglobulin (A2M) correlate with recovery from sepsis and 2) STAT-3 activation and A2M transcription reflect intrahepatic and not serum IL-6. Nonlethal sepsis was induced in rats by single puncture cecal ligation and puncture (CLP) and lethal sepsis via double-puncture CLP. STAT-3 activation and A2M transcription were detected at 3-72 h and intrahepatic IL-6 at 24-72 h following single-puncture CLP. All were detected only at 3-16 h following double-puncture CLP and at lower levels than following single-puncture CLP. Loss of serum and intrahepatic IL-6 activity after double-puncture CLP correlated with mortality. Neither intrahepatic nor serum IL-6 levels correlated with intrahepatic IL-6 activity. STAT-3 activation following single-puncture CLP inversely correlated with altered transcription of gluconeogenic, ketogenic, and ureagenic genes. IL-6 may have both beneficial and detrimental effects in sepsis. Fulminant sepsis may decrease the ability of hepatocytes to respond to IL-6.

Multiple versus Single Rubber Band Ligation for Internal Hemorrhoids: A Review of 450 Consecutive Cases

In the last 10 years we have treated 450 consecutive patients symptomatically affected by 1st- or 2nd-degree hemorrhoids with rubber band ligation. The patients were divided into 2 groups: group A, 216 patients treated from 1983 to 1989, and group B, 234 patients treated from 1990 to 1993. In group A multiple ligations (2 or 3) were made in a single session. In the group B one ligation at a time was performed. In group A satisfactory results were obtained in 78% of the cases, whereas in group B the results were good in 91% of patients (p < 0.01). Complications also decreased: bleeding decreased from 25 to 5% and pain from 22 to 9% (p < 0.01). Sepsis was present in 30% of the group-A cases, but was never seen in group B. In conclusion rubber band ligation offers better results if it is made with one ligation at a time, repeating the operation every 4 weeks.