BILIARY HYPERPRESSURE IN RAT EXTRAHEPATIC CHOLESTASIS ALTERS HORSERADISH PEROXIDASE BILIARY EXCRETION

1. The authors investigated the effect of two extrahepatic cholestasis models (one by bile duct ligation and the other by choledocho-jugular fistula) on the hepatic clearance of horseradish peroxidase in male Sprague-Dawley rats divided into four groups. 2. In groups A (n = 5 rats) and B (n = 5), bile duct ligation was performed, while a choledocho-jugular fistula was created in groups C (n = 5) and D (n= 7). A 10 mg intravenous bolus of horseradish peroxidase was injected after 24 h (groups A and C), 48 h (groups B and D) or 1 h (Group E; five sham-operated rats). Serum and bile samples were then serially collected for 2 h. 3. In all groups, serum horseradish peroxidase levels increased soon after injection and then rapidly decreased, the curves being similar. Biliary excretion increased for 30 min and then slowly decreased. The highest horseradish peroxidase biliary concentrations and outputs were found in Group B followed by Group A; both groups had significantly higher levels than Group E. No difference was found between horseradish peroxidase biliary excretion of groups C and D and that of sham-operated rats. 4. When each group was considered separately, sampling times correlated with the corresponding ratios of bile/ plasma HRP. Significant differences were found between the relative slopes of groups A, B and E, but not between those of groups C, D and E. 5. In conclusion, bile duct obstruction greatly affects the plasma-bile transfer of fluid phase markers, such as horseradish peroxidase, while single retention, caused by choledocho-jugular fistula, has no influence. The increased biliary hyperpressure related to the duration of cholestasis may account for the degree of horseradish peroxidase transfer which, in turn, probably depends on an enhanced paracellular passage.     

Prospective randomized multicentre study comparing stapler haemorrhoidopexy with Doppler‐guided transanal haemorrhoid dearterialization for third‐degree haemorrhoids

Aim Doppler‐guided transanal haemorrhoid dearterialization (THD) and stapler haemorrhoidopexy (SH) have been demonstrated to be less painful than the Milligan–Morgan procedure. The aim of this study was to compare the effectiveness of THD vs SH in the treatment of third‐degree haemorrhoids in an equivalent trial. Method One hundred and sixty‐nine patients with third‐degree haemorrhoids were randomized online to receive THD (n = 85) or SH (n = 84) in 10 Colorectal Units in which the staff were well trained in both techniques. The mean follow‐up period was 17 (range 15–20) months. Results Early minor postoperative complications occurred in 30.6% of patients in the THD group and in 32.1% of patients in the SH group. Milder spontaneous pain and pain on defecation were reported in the THD group in the first postoperative week, but this was not statistically significant. Late complications were significantly higher (P = 0.028) in the SH group. Residual haemorrhoids persisted in 12 patients in the THD group and in six patients in the SH group (P = 0.14). Six patients in the SH group and 10 in the THD group underwent further treatment of haemorrhoids (P = 0.34). No differences were found in postoperative incontinence. The obstructed defecation score (ODS) was significantly higher in the SH group (P < 0.02). Improvement in quality of life was similar in both groups. Postoperative in‐hospital stay was 1.14 days in the THD group and 1.31 days in the SH group (P = 0.03). Conclusion Both THD and SH techniques are effective for the treatment of third‐degree haemorrhoids in the medium term. THD has a better cost‐effective ratio and lower (not significant) pain compared with SH. Postoperative pain and recurrence did not differ significantly between the two groups.     

How to report the antinuclear antibodies (anti-cell antibodies) test on HEp-2 cells: guidelines from the ICAP initiative

Immunologic Research - Results of the anti-nuclear antibodies-indirect immunofluorescence assay (anti-cell antibodies test) on HEp-2 cell substrates should be communicated to clinicians in a...     

Femoral nerve damage after abdominal rectopexy

Iatrogenic femoral nerve damage has already been described after hysterectomy, but never after abdominal rectopexy. We report the occurrence of femoral nerve injury in six of twenty-four patients operated on for complete rectal prolapse (n=21) or rectorectal intussusception (n=3). Four patients had unilateral and two bilateral lesions. All six patients had clinical and electromyographic (EMG) assessment. EMG findings were given a score from 0 (complete denervation) to 5 (normal findings). During the immediate postoperative period all patients complained of reduced cutaneous sensation of the anterior surface of the thigh and knee, and quadriceps weakness. EMG showed complete denervation in one patient, marked denervation in three, and slight or moderate denervation in the remaining two. In five patients there was complete clinical resolution at 3 to 12 months postoperatively, while one showed an improvement only. EMG control performed in four patients showed a full recovery in three. Two patients refused this examination. We believe femoral nerve damage was caused by the large-bladed self-retaining retractors used, which directly or indirectly compressed the femoral nerve.

---

Résumé Une lésion iatrogénique du nerf fémoral a déjà été décrite après hystérectomie mais n'a jamais été rapportée après rectopexie abdominale. Nous rapportons 6 cas d'atteinte du nerf fémoral survenus dans un collectif de 24 patients dont 21 ont été opérés d'un prolapsus rectal complet et dont 3 ont été opérés d'une intussusception. Quatre patients présentaient des lésions unilatérales et 2 des lésions bilatérales. Les 6 patients ont été investigués cliniquement et par électromyographie. L'EMG a permis d'établir un score allant de 0 (dénervation complète) à 5 (constatations normales). Durant la période postopératoire immédiate tous les patients se sont plaints d'une diminution de la perception tactile sur la surface antérieure de la cuisse et du genou ainsi que d'une faiblesse du quadriceps. L'EMG a montré une dénervation complète chez un patient, une dénervation importante chez 3 malades et une dénervation modérée ou réduite chez 2 patients. Chez 5 patients la symptomatologie s'est normalisée en 3 à 12 mois en postopératoire alors qu'un malade ne présentait qu'une amélioration. Un EMG de contrôle réalisé chez 4 patients a montré une normalisation complète chez 3 d'entre eux. 2 malades ont refusé un EMG de contrôle. Nous pensons que l'atteinte du nerf fémoral résulte de l'emploi d'une large lame d'écarteur orthostatique qui comprime directement ou indirectement le nerf fémoral.

     

Assessment of humoral and cellular immunity induced by the BNT162b2 SARS-CoV-2 vaccine in healthcare workers,elderly people,and immunosuppressed patients with autoimmune disease

The development of vaccines to prevent SARS-CoV-2 infection has mainly relied on the induction of neutralizing antibodies (nAbs) to the Spike protein of SARS-CoV-2, but there is growing evidence that T cell immune response can contribute to protection as well. In this study, an anti-receptor binding domain (RBD) antibody assay and an INFγ-release assay (IGRA) were used to detect humoral and cellular responses to the Pfizer-BioNTech BNT162b2 vaccine in three separate cohorts of COVID-19-naïve patients: 108 healthcare workers (HCWs), 15 elderly people, and 5 autoimmune patients treated with immunosuppressive agents. After the second dose of vaccine, the mean values of anti-RBD antibodies (Abs) and INFγ were 123.33 U/mL (range 27.55–464) and 1513 mIU/mL (range 145–2500) in HCWs and 210.7 U/mL (range 3–500) and 1167 mIU/mL (range 83–2500) in elderly people. No correlations between age and immune status were observed. On the contrary, a weak but significant positive correlation was found between INFγ and anti-RBD Abs values (rho = 0.354, p = 0.003). As to the autoimmune cohort, anti-RBD Abs were not detected in the two patients with absent peripheral CD19⁺B cells, despite high INFγ levels being observed in all 5 patients after vaccination. Even though the clinical relevance of T cell response has not yet been established as a correlate of vaccine-induced protection, IGRA testing has showed optimal sensitivity and specificity to define vaccine responders, even in patients lacking a cognate antibody response to the vaccine.

     

Syk is a dual-specificity kinase that self-regulates the signal output from the B-cell antigen receptor

Upon B-cell activation, the signaling subunits Ig-α and Ig-β of the B-cell antigen receptor become phosphorylated not only on tyrosines but also on serine residues. Using a specific antibody, we show that serine 197 (S197) in the cytoplasmic tail of Ig-α is phosphorylated upon B-cell antigen receptor activation, and that this modification inhibits the signal output of the B-cell antigen receptor. Surprisingly, we found that the well-known protein tyrosine kinase Syk (spleen tyrosine kinase) phosphorylates S197 on Ig-α, thus not only activating but also inhibiting signaling from the B-cell antigen receptor. This finding identifies Syk as a dual-specificity kinase and establishes a previously unexplored paradigm for the self-regulation of biological signaling processes.     

Analytical variability in the determination of anti-double-stranded DNA antibodies: the strong need of a better definition of the old and new tests

Anti-dsDNA antibodies are a heterogeneous group of antibodies, quite specific for SLE. Their variability is related to the assay used, the immunoglobulin class secondary antibody, and the dsDNA source. The standardization of measuring anti-dsDNA antibodies is still poor and different methods yield different results. Several novel technologies were developed during the last decades that represent viable alternatives to the traditional methods such as the chemiluminescent immunoassay (CIA) and multiplex flow immunoassay (MFI). Additionally, positive results for anti-dsDNA antibodies can be detected in patients with inflammatory arthritis (IA) treated with different biologics reducing its clinical specificity for SLE. Anti-dsDNA antibody levels were evaluated in 246 patient samples: 70 SLE and 176 disease control (including 96 IA during treatment with different biologics), using three enzyme immunoassays (indirect enzyme immunoassay, Bio-Rad Laboratories; chemiluminescent immunoassay, Inova Diagnostics; multiplex flow immunoassay, Bio-Rad Laboratories) and three Crithidia luciliae immunofluorescence tests (CLIFT) (Euroimmun AG, Bio-Rad Laboratories, INOVA Diagnostics). Diagnostic performances were assessed both including and excluding the IA patients. Agreements, measured by the Cohen’s Kappa between all methods, ranged from moderate to substantial (0.47–0.68). The clinical sensitivities for the anti-dsDNA antibody tests varied from 5.7% by CLIFT A up to 33.3% provided by EIA while the clinical specificities varied from 89.8% by MFI to 98.9% provided by CLIFT B and C. Newer technologies, such as MFI and CIA, showed great potential as a diagnostic application. Significant variations among anti-dsDNA antibody assays were observed confirming the lack of standardization.     

Hot or cold in anal pain?

In 26 volunteers without anorectal complaints, and in 31 patients with anorectal problems such as hemorrhoidal disease, anal fissure, and proctalgia fugax, baseline resting anal canal pressures were recorded manometrically for 5 minutes at room temperature (23° C). In 16 volunteers (Group A) and 21 patients (group B) anorectal manometry was then performed while the anus was immersed in water at varying temperatures (5° C, 23° C, and 40° C). In ten volunteers (Group A′) and ten patients (Group B′) resting pressures were recorded for an additional 30 minutes following immersion for 5 minutes at 40° C. In all subjects (at leastP<0.01), resting anal canal pressures diminished significantly from baseline after immersion at 40° C, but remained unchanged in all subjects after immersion at 5° C and 23° C. In Group A′, anal canal pressures remained significantly reduced for 15 minutes (P<0.02). In Group B′, significant reduction in resting pressure lasted 30 minutes (P<0.02). Wet heat applied to the anal sphincter apparatus significantly and reproducibly decreased resting anal canal pressures over time, and therefore was likely to benefit patients after anorectal operations and those with anorectal pain.     

The relaxing effect of caerulein on isolated human internal anal sphincter

In the attempt to find a pharmacological treatment for the spasm of the internal anal sphincter, usually associated with anal fissures, the activity of caerulein on human internal and sphincter was investigated in vitro and in vivo. In the isolated distal part of the internal and sphincter, caerulein (0.61 microM) depressed resting muscle tone and caused marked relaxation of norepinephrine-contracted preparations. The effect of caerulein was reduced by atropine and increased by physostigmine, suggesting that it was largely due to the release of acetylcholine. In vivo, intravenous infusion of caerulein, both to healthy volunteers and to subjects affected by anal fissures and anal sphincter hypertone, did not modify the values of internal anal sphincter pressure. The lack of spasmolytic effect of caerulein in vivo may have been due to the relatively unimportant influence of cholinergic neurons on the control of internal anal sphincter tone. Alternatively, the presence of fibrosis caused by anal fissures could hinder sphincter relaxation.