Transfer of the first intercostal nerve to supra- and infraspinatus muscles: an anatomical study and report of the first case.

We have assessed the anatomical feasibility of a transfer of the first intercostal nerve to the supra- and infraspinatus muscles and report on the first clinical application. Ten fresh cadavers were dissected for this study. Histomorphometric analysis showed the fascicular surface area of the first intercostal nerve at its origin (0.38 mm(2)) to be comparable to the suprascapular nerve (0.81 mm(2)). The first intercostal nerve is usually a pure motor nerve. Preservation of the spinal accessory nerve, lack of donor site morbidity and direct suture without nerve graft are the other advantages of this transfer. Its principal indication is in lesions of the upper brachial plexus, used in association with neurotisation of two other intercostal nerves to the anterior branch of the axillary nerve. At 21 months follow-up there was useful motor reinnervation in the first clinical case.

Increased expression of MDR1 mRNAs and P-glycoprotein in placentas from HIV-1 infected women.

P-glycoprotein transports several compounds including protease inhibitors, actually used in the clinical management of HIV-1 infection. Since P-glycoprotein is expressed in placental trophoblasts, its efflux activity could interfere with placental transfer of antiretrovirals. The purpose of this study was to investigate the expression of P-gp-encoding MDR1 gene and P-gp itself in full-term placentas from uninfected (n=35) and HIV-1 infected women (n=24). MDR1 transcripts were quantified by real-time PCR using relative (MDR1 normalized upon 28S levels) and absolute (copy number) determinations. P-glycoprotein localization and expression were evaluated by immunohistochemistry and western blot analysis, respectively. Relative or absolute PCR quantification showed a significant 3.3-fold (p<0.0009) or 3.7-fold (p<0.0002) mean increase in MDR1 placental transcription in HIV-infected compared to non-infected women, respectively. Ratios of individual HIV-positive values to HIV-negative mean ranged from 0.1 to 21.8. Moreover a significant 2.5-fold increased expression of immunoreactive P-glycoprotein was evidenced in placentas from HIV-infected women (p<0.0001). This MDR1 overexpression was observed in a similar extent in placentas from pregnant women treated with Zidovudine alone or in combination with Nelfinavir and/or Lamivudine. Our findings suggest that P-glycoprotein in placentas from HIV-infected women would contribute to modulate the materno-fetal transport of antiretrovirals across the placental barrier and consequently diminish fetal exposure to these compounds.

Morphometric study of the cervical course of the hypoglossal nerve and its application to hypoglossal facial anastomosis.

The hypoglossal nerve is used classically in salvage of facial paralyses in the absence of spontaneous recovery. A variety of ways of transferring and suturing the hypoglossal nerve to the distal segment of the facial nerve have been reported. In order to determine which mode of reconstruction is the best for neurotisation of the facial nerve, the caliber of the hypoglossal nerve was studied in ten subjects at the level of proximal and distal parts of the trunk and the cervical loop. The fascicular surface area of the cervical branch is inadequate for use. The distal extremity of the hypoglossal nerve has an ideal caliber to be sutured to the facial nerve trunk and the proximal part is large enough to allow partial harvesting of the hypoglossal nerve for neurotisation of the facial nerve.

Nerve repair using a composite graft of vein and denatured skeletal muscle: morphologic analysis.

Five groups of six rats each underwent sciatic resection, bridged with a 20-mm-long graft. Five different types of graft were used: (1). vein; (2). fresh muscle; (3). denatured muscle; (4). vein and fresh muscle; and (5). vein and denatured muscle. Denaturation was obtained by dry thermal shock in a microwave oven. Morphologic analysis took place at 6 months. The macroscopic appearance of the graft was continuous in all type 4 and 5 animals. Morphometric analysis within the graft revealed a significantly greater number of fibers in type 3 animals. Average fiber diameter and g ratio were higher, although not significantly, in type 5 animals. Distal to the graft, average fiber diameter and g ratio were significantly higher in type 5 rats. Denaturation by exposure to thermal shock significantly improves axonal regeneration, both quantitatively and qualitatively. The regeneration rate seems to be more rapid within denatured muscle. Denaturing muscle by exposure to thermal shock provides an excellent guide for axonal regrowth, but axonal leaks may occur. This problem is solved by a muscle-in-vein graft technique.

Endoluminal smooth muscle cell seeding limits intimal hyperplasia.

PURPOSE: Intimal hyperplasia is one of the main responses of the vascular wall to injury. In the current study, we tested the hypothesis that endoluminal seeding of host syngeneic vascular cells could limit intimal hyperplasia induced by either mechanical deendothelialization or chronic allograft rejection in rat aorta. METHODS: An experimental model of in situ seeding of syngeneic endothelial cells, smooth muscle cells (SMCs), and fibroblasts (FIBs) was used in mechanically deendothelialized and allografted aortas. In a preliminary study, the ability of the three cell types (n = 5 per group) to seed on the deendothelialized luminal surface of the aortic wall was evaluated after 2 days, with the use of fluorescent PKH as marker. In the first model, the abdominal aorta of Lewis rats was deendothelialized (n = 6) or deendothelialized and seeded with either SMCs (n = 6) or FIBs (n = 6) before flow was restored. In the allograft model, aortas were harvested from dark agouti rats and orthotopically grafted in Lewis receivers, directly (n = 6) or after deendothelialization. Deendothelialization was performed alone (n = 6) or associated with the seeding of similar host (Lewis) syngeneic SMCs (n = 6) or FIBs (n = 6). Results were evaluated at 2 months with histologic and morphometric methods. RESULTS: SMCs and FIBs were able to adhere in situ to the deendothelialized aortic wall, whereas endothelial cells were not. In mechanically deendothelialized aortas, the seeding of syngeneic SMCs led to a significant reduction in intimal thickness compared with deendothelialized aortas or FIB-seeded aortas (26.9 +/- 1.7 microm vs 55.5 +/- 1.7 and 56.7 +/- 1.7 microm, respectively), and a lower nuclear content (382.2 +/- 35.7 microm(2) vs 779.6 +/- 65.9 and 529.6 +/- 24.3 microm(2), respectively) of neointima. After SMC seeding, intimal hyperplasia was richer in elastin, whereas after FIB seeding it was richer in collagen. In allografts, the seeding of syngeneic SMC led to a significant reduction in intimal thickness compared with control aortas, deendothelialized aortas, or FIB-seeded aortas (31.6 +/- 1.1 microm vs 88.55 +/- 2.8, 74.6 +/- 2.9, and 85.7 +/- 2.6 microm, respectively), and a reduced nuclear content of the neointima (444.9 +/- 23.4 microm(2) vs 1529.1 +/- 116, 972.3 +/- 50, and 645.2 +/- 32.4 microm(2), respectively). Differences observed in the extracellular matrix composition were equivalent to those observed in the mechanically deendothelialized model. CONCLUSIONS: Our results suggest that endoluminal seeding of syngeneic SMCs can be effective in reducing intimal hyperplasia both in a deendothelialization model and in arterial allografts. SMC and FIB endoluminal seeding led to a significatively different accumulation of extracellular matrix in the intima.

Cytogenetic analysis of trophoblasts by comparative genomic hybridization in embryo-fetal development anomalies.

Cytogenetic studies of spontaneous abortions or intrauterine fetal death depend on conventional tissue culturing and karyotyping. This technique has limitations such as culture failure and selective growth of maternal cells. Fluorescent in situ hybridization (FISH) using specific probes permits diagnosis of aneuploidies but is limited to one or a few chromosomal regions. Comparative genomic hybridization (CGH) provides an overview of chromosomal gains and losses in a single hybridization directly from DNA samples. In a prospective study, we analyzed by CGH trophoblast cells from 21 fetuses in cases of spontaneous abortions, intrauterine fetal death or polymalformed syndrome. Six numerical chromosomal abnormalities including one trisomy 7, one trisomy 10, three trisomies 18, one trisomy 21 and one monosomy X have been correctly identified by CGH. One structural abnormality of the long arm of chromosome 1 has been characterized by CGH. One triploidy and two balanced pericentromeric inversions of chromosome 9 have not been identified by CGH. Sexual chromosomal constitutions were concordant by both classical cytogenetic technique and CGH. Contribution of trophoblast analysis by CGH in embryo-fetal development anomalies is discussed.

Incisive suture (fissure) in the human fetus: radiographic and histologic study.

OBJECTIVE: An anatomic study, in man, of the structure commonly known as the incisive suture (sutura incisiva) or incisive fissure has been performed to determine whether this structure belongs to the morphofunctional concept of a facial suture. MATERIAL: Eighteen palates of human fetuses of 9 weeks to 29 weeks after conception. METHOD: Anatomic, radiographic examination of the palate. Histologic study of sagittal and parasagittal sections of the palates (3 mm each). RESULTS: The histologic aspect is that of a facial suture, with very poor vascularization. This suture is partial, limited laterally by osseous trabeculae. CONCLUSIONS: This particular suture, whose function in growth of the palatal process of the maxilla is discussed, could represent the phylogenetic vestige of the incisive-maxillary suture present in all nonhuman mammals.

[The anatomic lobulation of the prostate, a controversial description]. / La lobulation anatomique de la prostate, une description controversée.

Vésalius, in 1543, described, for the first time, the prostate as an unique organ. But, in the 19th century, two schools confronted; for Cruveilhier and Testut, the prostate was made of several lobes, when Cloquet and Sappey thought it as a unique zone. Albarran, in 1902, described the sub-uretral glands. Thereafter, Cuneo, in 1911 and Franks, in 1954, described two zones, one, internal, formed by the Albarran's glands, and the other, external, concerning the whole prostatic gland. On the contrary, Lowsley, in 1912, and Gil Vernet, in 1953, described several lobes, 5 for Lowsley, 3 for Gil Vernet. Recently, in 1968, and 1978, McNeal had made the proof that the prostate is histologically and anatomically heterogeneous, with three zones, transitional, central and peripheral ones.

Reduced number of Langerhans cells in oesophageal mucosa from AIDS patients.

AIMS: The oesophageal mucosa is a frequent target of opportunistic infections in human immunodeficiency virus (HIV) infection. Langerhans cells (LC) are known as a target and reservoir of HIV in the skin. The aim of this study was to characterize oesophageal LC in HIV-infected patients. METHODS AND RESULTS: Thirty oesophageal biopsies were obtained from 29 patients (median age 35.5), all in stage IV of the HIV Center of Disease Control Classification. We performed histological assessment of the oesophageal mucosa and immunohistochemical detection of oesophageal LC using an anti-CD1a antibody, followed by morphometric analysis. Biopsies from 17 noninfected patients were studied using the same procedure. LC in oesophageal mucosa of the HIV positive patients showed a significantly and dramatically decreased number (LC(N) median = 5.85/mm2) and surface/epithelial surface (LC (S) ratio = 0.09) when compared with HIV-negative controls (LC(N) median = 29.7/mm2, LC(S) ratio = 1.83) with P = 0.003 for LC(N) and P < 0.0001 for LC(S). CONCLUSION: These data suggest that oesophageal LC are, like their epidermal counterparts, a preferential target for HIV infection. Their alterations may provide a clue to the pathogenesis of the decreased local oesophageal immunity and to the occurrence of opportunistic infections.

Morphometric study of the upper intercostal nerves: practical application for neurotizations in traumatic brachial plexus palsies.

UNLABELLED: The aim of the study was a morphometric evaluation of the intercostal nerves at different levels along their course in order to determine their adequacy in neurotizing the recipient nerves. The intercostal nerves were harvested from 5 cadavers. A biopsy of the nerve was obtained at 2 levels for each nerve in the parasternal region and at the level of the mid-axillary line. The musculocutaneous nerve was isolated at its origin from the lateral cord. Each harvested specimen was embedded in paraffin and sections were made using a microtome. These sections were then stained histochemically using HPS (Hematein, Phloxine, Safran). Real-time digitalisation of the video image under the microscope was performed. The sum of the different fascicular zones is the effective sensorimotor surface of the nerve at the level being studied. RESULTS: Direct suture of the upper three intercostal nerves to the musculocutaneous nerve is always possible upto the axillary fossa. The sixth intercostal nerve can be delivered upto this level in only 50% of cases without dissection of the musculocutaneous nerve upto its entry into the coracobrachialis. The musculocutaneous nerve presents a mean surface area of 2.64 mm2 while the nerve to the biceps has a mean surface area of 0.34 mm2 i.e. a ration of 1/8. The mean surface area of the intercostal nerves at the parasternal level is 0.23 mm2 while that at the axillary level is 0.34 mm2. Thus a loss of 33% in surface area occurs between the axillary and the parasternal levels. Our study confirms the insufficiency between the surface area of the intercostal nerves and the different nerve trunks to be neurotized. The relationship between the surface area of the musculocutaneous nerve and the three intercostal nerves is 26.72% with a minimum of 17.2%. If a fourth intercostal nerve is added, this ratio nerves appears to be a superior technique. We were able to deliver the sixth intercostal nerve for a direct suture to the musculocutaneous nerve in only half the cases.

Ticlopidine induced colitis: a histopathological study including apoptosis.

AIMS: To describe ticlopidine related microscopic colitis and to assess the occurrence of apoptosis in the colon epithelium. METHODS: A series of colorectal biopsy samples from nine patients with ticlopidine related chronic diarrhoea were analysed. Biopsies were also taken from five of these patients between two and four months after ticlopidine withdrawal. The number of apoptotic cells in the crypts/mm2 (apoptotic index) was calculated using in situ labelling by terminal deoxyribonucleotidyl transferase (TdT) mediated dUTP-biotin nick end labelling (TUNEL). All specimens were matched to normal colorectal specimens from a control group of comparable age and sex distribution. RESULTS: Histological examination of the colon biopsy specimens taken from all nine patients with ticlopidine related chronic diarrhoea showed characteristic features of microscopic colitis. The histology returned to normal when ticlopidine was withdrawn. Apoptotic cells were rarely found in controls, and the mean apoptotic index was 0.53. The apoptotic index was significantly higher (16.53) in ticlopidine related colitis, but decreased dramatically to control value when ticlopidine was withdrawn. CONCLUSION: Microscopic colitis can be induced by ticlopidine and is accompanied by an increase in epithelial apoptosis. Hence, increased apoptosis might be related to drug injury or might be part of microscopic colitis.

Detection and localization by in situ molecular biology techniques and immunohistochemistry of hepatitis C virus in livers of chronically infected patients.

Hepatitis C virus (HCV) detection in the livers of chronically infected patients remains a debatable issue. We used immunohistochemistry, in situ hybridization (ISH) alone or after microwave heating with FITC-labeled probes, RT-PCR with unlabeled primers followed by ISH (RT-PCR-ISH), and in situ RT-PCR with FITC-labeled primers (in situ RT-PCRd) to localize the virus in 38 liver biopsy specimens from 21 chronically infected HCV patients treated with interferon-alpha (IFN-alpha). Biopsies were taken at the beginning and end of IFN-alpha treatment and 1 year later. Results were compared with that of HCV-PCR in serum. RT-PCR-ISH and in situ RT-PCRd showed HCV signal in all liver biopsies even in responders with seronegative HCV PCR. This signal was intranuclear, diffuse, or peripheral, in hepatocytes, bile ductule cells, and lymphocytes. Cytoplasmic signals were occasionally observed. Whereas the percentage of labeled hepatocytes remained constant, the number of labeled lymphoid follicles decreased after INF-alpha therapy. Immunohistochemistry resulted in the same pattern of positivity but it was weaker and inconstant. This study indicates the persistency of HCV latency in IFN-alpha responders 1 year after IFN-alpha treatment cessation, a finding that certainly deserves confirmation.

Expression of integrins during liver organogenesis in humans.

Integrins play a pivotal role in organogenesis, by mediating the interactions between differentiating cells and the extracellular matrix. We analyzed the expression of integrins and their ligands during human liver organogenesis. The expression of beta1, beta3, and beta4 integrins and the distribution of several extracellular matrix proteins were studied by immunoperoxidase in fetal liver samples from 5 to 40 weeks' gestation. Hepatoblasts expressed only the beta1, alpha1, alpha5, alpha6, and alpha9 integrin chains. Fetal hepatocytes, emerging at the 8th week of gestation, initially retained the same combination of integrins, but presented a progressive decrease in their expression levels. After 15 weeks' gestation, the expression levels of beta1, alpha1, alpha5, and alpha9 reached levels comparable to those observed in the adult state. Alpha6 expression became undetectable after 30 weeks' gestation. As compared to hepatoblasts, intrahepatic biliary epithelial cells, differentiating at the 8th week of gestation in the ductal plate, were characterized by the progressive loss of alpha1, the marked induction of alpha6, and the de novo acquisition of the beta4, alpha2, and alpha3 integrin chains. The disappearance of integrin receptors for laminin on hepatocytes was associated with the rarefaction of laminin in the perisinusoidal matrix, whereas their induction on biliary epithelial cells was associated with laminin deposition at the point of contact with the ductal plate. In conclusion, integrins likely play an important role in the differentiation of the epithelial and endothelial cell populations of the liver.

Structural and functional differentiation of sinusoidal endothelial cells during liver organogenesis in humans.

During fetal life, human liver sinusoids, which differentiate between 4 and 12 weeks of gestation from capillaries of the septum transversum, must support an important hematopoietic function and acquire the structural and functional characteristics of adult sinusoids. To gain insight into their differentiation process, we studied the expression of (1) markers of continuous endothelia, absent from adult sinusoidal endothelial cells (PECAM-1, CD34, and 1F10); (2) functional markers of adult sinusoidal endothelial calls (CD4, 1CAM-1, CD32, and CD14); and (3) extracellular matrix components (laminin, tenascin, fibronectin, and thrombospondin) in 37 fetuses of different gestational ages. We identified two successive differentiation events. (1) An early structural differentiation, occurring from 5 to 12 weeks of gestation, was characterized by the loss of continuous endothelial cell markers and a reduction in the perisinusoidal amount of laminin and in the deposition of tenascin, fibronectin, and thrombospondin; at the end of this process, fetal liver sinusoids present structural characteristics comparable to those of the sinuses in adult hematopoietic bone marrow. (2) A later functional differentiation was characterized by the acquisition of the markers of adult sinusoidal endothelial cells, initiating at 10 weeks of gestation and completed by 20 weeks of gestation; this process likely contributes to adapt liver sinusoids to the specific functions of the adult hepatic tissue.

Effect of radiation therapy after radical prostatectomy on serum prostate-specific antigen measured by an ultrasensitive assay.

OBJECTIVES: To study prospectively the impact of adjuvant radiation therapy on the serum level of prostate-specific antigen (PSA), as measured by an ultrasensitive Yang Proscheck assay in patients with detectable serum PSA and a negative metastatic survey after radical prostatectomy for T1 or T2 prostate cancer. METHODS: Seventeen patients had a detectable serum PSA (2.40 +/- 2.1 ng/mL; range, 0.5 to 10) by the Yang polyclonal assay 2 to 71 months after radical prostatectomy for P2N0 (2 patients) or P3N0 (15 patients) prostate cancer. Metastatic workup (bone and computed tomography scan) was negative; 9 of 17 patients had a local recurrence documented by a positive biopsy of the vesicourethral anastomosis. All patients were treated by external radiotherapy, receiving 65 Gy on the prostate fossa over 5 weeks for an assumed low volume residual disease. Patients were followed up by determination of serum PSA every 3 months, using the Yang ultrasensitive assay for a mean duration of 14.4 months. RESULTS: In 17.6% of the patients (3 of 17) PSA became undetectable (less than 0.05 ng/mL) after radiotherapy. Radiotherapy had no impact on PSA in 35.3% (6 of 17). PSA decreased after radiation therapy within 6 months in 47.1% (8 of 17) and for up to 12 months in 2 patients, with a nadir of 0.28 ng/mL. All patients in this group experienced a secondary rise in PSA a mean of 10.6 months (range, 6 to 18 months) after radiotherapy. CONCLUSIONS: External radiotherapy has a limited impact on residual disease after radical prostatectomy, as assessed by its impact on PSA.

Clearance of serum PSA after open surgery for benign prostatic hypertrophy, radical cystectomy, and radical prostatectomy.

OBJECTIVE: To study the clearance of serum prostate-specific antigen (PSA) after several types of prostatic tissue ablation. METHODS: Serum PSA levels were measured (YANG Proscheck ultrasensitive assay) just before surgery, immediately after specimen removal, then twice weekly for 5 weeks or until it was undetectable (< 0.05 ng/ml) in patients undergoing radical cystoprostatectomy for bladder cancer (n = 10), or radical prostatectomy for T1 T2 prostate cancer (n = 18) and daily for 6 days after open surgery for benign prostatic hypertrophy (BPH) (n = 10). RESULTS: Open enucleation for BPH: the immediately postoperative PSA level was 6 times its preoperative value. It decreased following a monoexponential curve with a very short half-life of 0.55 +/- 0.39 days, range (0.14-1.3), reaching a value lower than the preoperative level in all cases, except one by day 3. After radical cystoprostatectomy: the decrease of serum PSA is monoexponential with a half life of 1.92 +/- 1.2 days (0.57-4.24) reaching undetectable level (< 0.05 ng/ml) in all patients by day 21. After radical prostatectomy: 11/18 patients (61%) showed a one-component exponential decrease in PSA with a half-life of 2.5 +/- 1.33 days (range 0.97-4.6 days), and 7/18 showed a two-component exponential decrease with a first half-life of 0.94 +/- 0.8 days and a second of 7.62 +/- 6.35 days); 100% of the patients reached undetectable serum PSA by day 28 in the first group compared to 14.2% of the patients with a two component exponential decrease (P < 0.01). There was no difference between these groups as far as preoperative PSA levels and specimen pathology were concerned. CONCLUSION: Serum clearance of PSA after extirpative prostatic surgery is closely related to the type and indication of procedure used. Radical cystoprostatectomy is probably the best model in which to study the pharmacokinetics of PSA.

[In situ gene amplification on tissue sections (in situ PCR). A new technique for pathologists]. / L'amplification génique in situ sur coupes tissulaires (PCR-in situ).

In situ polymerase chain reaction is a recent technique which combines the sensitivity of PCR reaction to intracellular localization of genomic sequences with the same specificity as in situ hybridization. This reaction is based on the in situ annealing and polymerisation of oligonucleotides complementary to nucleotides located at each side of the target DNA sequence to amplify. We describe the Hot Start PCR (DNA) and the Hot Start PCR after reverse transcription step (RNA). It allows to amplify some nucleic sequences to a high level, becoming easier to detect. The vizualisation can be realized by direct in situ PCR, the product obtained being directly identifiable by incorporation of labeled nucleotides or primers, or preferentially by indirect in situ PCR. In this case, the amplification is followed by in situ hybridization with labeled probes. This last procedure is more specific. Numerous controls are essential at each step of the technique for validating results.

Nerve transfer to biceps muscle using a part of ulnar nerve for C5-C6 avulsion of the brachial plexus: anatomical study and report of four cases.

Four patients with C5-C6 root avulsion after brachial plexus injury were treated with a transfer of part of a normal functioning nerve in the arm to the motor nerve of the biceps. Ten percent of the bulk of the ulnar nerve was harvested for a suture directly to the motor nerve of the biceps with no significant impairment of hand function.

Prostate-specific antigen density: a means to enhance detection of prostate cancer.

The ability of serum prostate-specific antigen (PSA) and PSA density (PSAD) to distinguish patients with prostate cancer from those with benign diseases of the prostate was assessed in 495 men. All men were evaluated with PSA determination, digital rectal examination (DRE), transrectal ultrasonography (TRUS) and ultrasound-guided prostatic biopsies. PSA was analysed by the polyclonal (Yang) assay. Prostate volume was estimated from TRUS. PSAD was determined by dividing the serum PSA by the volume of the prostate. Prostatic biopsies identified cancer in 246 of the 495 patients (49.7%). The entire group was divided into 6 subgroups according to PSA level at presentation. Cancer and noncancer patients were compared in each subgroup with respect to the values of PSA, prostate volume and PSAD. For the entire group of patients, there was no statistically significant advantage, for PSAD over serum PSA alone, in distinguishing between benign and malignant prostatic conditions. However, when patients were stratified according to PSA level, PSAD was statistically significantly superior to serum PSA alone in the detection of prostate cancer for PSA values in the intermediate range (2.6-30 ng/ml). This analysis with respect to the DRE and TRUS results showed PSAD to be superior to PSA when both examinations are normal. Our results demonstrate that the influence of PSAD level on cancer detection proportionally increases as the PSAD value increases. Curves constructed from the incidence of prostate cancer according to PSAD values may be useful to select patients with intermediate levels of serum PSA, and normal DRE and TRUS for prostatic biopsies.