Effect of human albumin on TCD vasospasm, DCI, and cerebral infarction in subarachnoid hemorrhage: the ALISAH study.

BACKGROUND AND PURPOSE: The neuroprotective effects of human albumin have been studied in animal models of stroke and in humans with various intracranial disorders. We investigated the effect of 25 % human albumin (ALB) on mean cerebral blood flow velocities (MCBFV), delayed cerebral ischemia (DCI), and cerebral infarction. METHODS: We studied patients from the Albumin in Subarachnoid Hemorrhage (ALISAH) pilot clinical trial. We collected data on MCBFV as measured by transcranial Doppler ultrasound (TCD), incidence of DCI, and cerebral infarctions on head computed tomography (CT) scan at 90 days. RESULTS: TCD showed vasospasm in 75 % (n = 15), 55 % (n = 11), and 29 % (n = 2) of subjects in dosage tiers 1, 2, and 3, respectively. DCI was present in 20 % (n = 4), 15 % (n = 3), and 14 % (n = 1) of subjects in dosage tiers 1, 2, and 3, respectively. Cerebral infarctions were seen in 45 % (5 of 9), 27 % (3 of 18), and 25 % (1 of 4) of subjects who had follow-up head CT scans in dosage tiers 1, 2, and 3, respectively. CONCLUSIONS: Higher dosages of ALB were associated with a lower incidence of TCD vasospasm, DCI, and cerebral infarction at 90 days in a dose-dependent manner.

Reduced meiotic recombination on the XY bivalent is correlated with an increased incidence of sex chromosome aneuploidy in men with non-obstructive azoospermia.

Both aberrant meiotic recombination and an increased frequency of sperm aneuploidy have been observed in infertile men. However, this association has not been demonstrated within individual men. The purpose of this study was to determine the association between the frequency of recombination observed in pachytene spermatocytes and the frequency of aneuploidy in sperm from the same infertile men. Testicular tissue from seven men with non-obstructive azoospermia (NOA) and six men undergoing vasectomy reversal (controls) underwent meiotic analysis. Recombination sites were recorded for individual chromosomes. Testicular and ejaculated sperm from NOA patients and controls, respectively, were tested for aneuploidy frequencies for chromosomes 9, 21, X and Y. There was a significant increase in the frequency of pachytene cells with at least one achiasmate bivalent in infertile men (12.4%) compared with controls (4.2%, P = 0.02). Infertile men also had a significantly higher frequency of sperm disomy than controls for chromosomes 21 (1.0% versus 0.24%, P = 0.001), XX (0.16% versus 0.03%, P = 0.004) and YY (0.12% versus 0.03%, P = 0.04). There was a significant correlation between meiotic cells with zero MLH1 foci in the sex body and total sex chromosome disomy (XX + YY + XY) in sperm from men with NOA (r = 0.79, P = 0.036).

Sperm aneuploidy frequencies analysed before and after chemotherapy in testicular cancer and Hodgkin's lymphoma patients.

BACKGROUND Multicolour fluorescent in situ hybridization was utilized to detect sperm aneuploidy for chromosomes 13, 21, X and Y in testicular cancer and Hodgkin's lymphoma chemotherapy patients. METHODS Aneuploidy was assessed before, and 6, 12 and/or 18-24 months after, the initiation of chemotherapy, and compared with age matched controls. 635 396 sperm were scored blindly with 5000 sperm/patient/chromosome/ time point, where sperm was available. (First two phrases have been reversed). RESULTS Comparing testicular cancer and Hodgkin's lymphoma patients to each other and with controls, cancer-specific differences were identified. Hodgkin's lymphoma patients, particularly, exhibited significantly increased aneuploidy frequencies for all chromosomes throughout treatment. At 6 months, all cancer patients showed significantly increased frequencies of XY disomy and nullisomy for chromosomes 13 and 21. In general, aneuploidy frequencies declined to pretreatment levels 18 months after treatment initiation, but increased aneuploidy frequencies persisted in some chromosomes for up to 24 months. CONCLUSIONS Because of elevated aneuploidy frequencies prior to and up to 24 months from the start of chemotherapy, patients should receive genetic counselling about the potentially increased risk of an aneuploid conceptus from sperm cryopreserved prior to chemotherapy, and for conceptions up to 2 years after the initiation of treatment.

The relationship between meiotic recombination in human spermatocytes and aneuploidy in sperm.

BACKGROUND: We have previously demonstrated that a decreased recombination frequency between human X and Y chromosomes is associated with the production of aneuploid 24,XY sperm. This study's aim was to determine the relationship between recombination frequency in human pachytene spermatocytes and aneuploidy frequencies in individual chromosomes in sperm from the same men. METHODS: Six previously fertile vasectomy reversal patients donated testicular tissue for meiotic analysis of pachytene spermatocytes using immunocytogenetic techniques for visualization of the synaptonemal complex and recombination sites (MLH1). Individual meiotic chromosomes were identified with centromere-specific multicolor fluorescence in situ hybridization (FISH), and the number of MLH1 signals was recorded for individual chromosomes. An ejaculated sperm sample was obtained from each patient 2-26 months post-reversal for FISH analysis of sperm aneuploidy frequencies of chromosomes 1, 9, 13, 21, X and Y. RESULTS: There was no significant correlation between meiotic recombination frequency and sperm aneuploidy for any individual chromosome. Similarly, there was no correlation between aneuploid sperm and bivalents with no recombination. CONCLUSIONS: The study provides unique data on intra-individual human recombination and aneuploidy events. It also demonstrated for the first time that men do not have an increased frequency of sperm aneuploidy 5-9 years post-vasectomy.

Atrial fibrillation in the intact unanesthetized dog: hemodynamic effects during rest, exercise, and beta-adrenergic blockade.

The effects of atrial fibrillation were studied in 12 healthy unanesthetized dogs, 9 to 49 days after surgical implantation of transducers for measurement of aortic flow and left ventricular diameter. Atrial fibrillation and pacing at comparable ventricular rates were induced by electrical stimulation of the right atrial appendage, and their effects were compared with observations made during sinus rhythm in each dog. At rest, cardiac output and mean arterial pressure were not significantly different during sinus rhythm, atrial fibrillation, and atrial pacing. After beta-adrenergic blockade with propranolol, cardiac output during fibrillation was significantly less than that during pacing at comparable ventricular rates. Arterial pressure was not detectably altered. During moderately severe treadmill exercise by six dogs, cardiac output fell significantly upon induction of fibrillation. After pentobarbital anesthesia fibrillation caused decrements in cardiac output and arterial pressure that were accentuated after thoracotomy.These observations suggest the existence of compensatory mechanisms that maintain an essentially constant cardiac output when atrial fibrillation is induced in healthy unanesthetized dogs at rest. These mechanisms appear to fail during moderately severe exercise, beta-adrenergic blockade, and pentobarbital anesthesia.

Discontinuities and unsynapsed regions in meiotic chromosomes have a trans effect on meiotic recombination of some chromosomes in human males.

During meiosis, homologous chromosome pairing and synapsis are essential for subsequent meiotic recombination (crossing-over). Discontinuous regions (gaps) and unsynapsed regions (splits) were most frequently observed in the heterochromatic regions of bivalent synaptonemal complex (SC) 9, and we have previously demonstrated that gaps and splits significantly altered the distribution of MLH1 recombination foci on SC 9. Here, immunofluorescence techniques (using antibodies against SC proteins and the crossover-associated MLH1 protein) were combined with a centromere-specific fluorescence in situ hybridization technique that allows identification of every individual chromosome. The effect of gaps/splits on meiotic recombination patterns in autosomes other than chromosome 9 during the pachytene stage of meiotic prophase was then examined in 6,026 bivalents from 262 pachytene cells from three human males. In 64 analyzed cells with a gapped SC 9, the frequency of MLH1 foci in SCs 5 and 10 and in SC arms 10q, 11p and 16q was decreased compared to 168 analyzed cells with a normally-synapsed SC 9 (controls). In 24 analyzed cells with splits in SC 9, there was a significant reduction in MLH1 focus frequency for SC 5q and the whole SC5 bivalent. The positioning of MLH1 foci on other SCs in cells with gapped/split SC 9 was not altered. These studies suggest that gaps and splits not only have a cis effect, but may also have a trans effect on meiotic recombination in humans.

Analysis of replication protein A (RPA) in human spermatogenesis.

Replication protein A (RPA) has been identified as a component of early recombination nodules. It is thought to stimulate homologous pairing and strand exchange reactions. The expression pattern of RPA in human spermatocytes has been analysed using immunocytogenetic techniques on testicular biopsies from adult male patients. What appears to be connecting RPA-filaments was observed between as yet unsynapsed homologous regions at early stages of zygotene. RPA foci were also observed in synaptic segments at zygotene and early pachytene, in numbers that peak at the end of zygotene. The presence of a localization pattern for RPA was also detected, but statistical analysis of distances between adjacent RPA foci shows that this pattern does not always follow a gamma distribution. Finally, it was determined that RPA is absent from non-centromeric heterochromatin in chromosome 9. The observed bridge-like structure could be the visualization of a proposed pre-synaptic RPA role in the strand invasion that precedes the formation of a Holliday Junction. These observations strengthen the original pre-synaptic model, although the visualization of post-synaptic RPA foci may indicate the presence of a different role for this protein during homologous recombination.

Studying meiosis: a review of FISH and M-FISH techniques used in the analysis of meiotic processes in humans.

It is well known that chromosome in situ hybridization allows the unequivocal identification of targeted human somatic chromosomes. Different fluorescent in situ hybridization (FISH) techniques have been developed throughout the years and, following the mitotic studies, meiotic analyses have been performed using these different techniques. The introduction of M-FISH techniques to the analysis of meiotic cells has allowed the study of meiotic processes for every individual human chromosome. In this paper, we review the different FISH and M-FISH techniques that have been used on human meiotic cells in both men and women.

Imaging of human melanoma xenografts in nude mice with a radiolabeled monoclonal antibody.

External photoscanning with display of radioactivity data as a color-scaled image detected xenografts of human melanoma in male nude inbred mice of BALB/c background 48 hours after injection of 131I-labeled monoclonal IgG 225.28S that is specific for human melanoma. A 131I-labeled polyclonal goat IgG against human melanoma-associated antigens could also image the tumor, but with this preparation there was considerable localization of radioactivity in normal tissues, resulting in less satisfactory tumor definition. Labeled normal mouse IgG did not image the melanoma grafts. Assay of radioactivity in tissues of melanoma-grafted mice confirmed tumor-specific localization of the antimelanoma antibodies. The tumor:blood ratio of radioactivity was 6.55 with the monoclonal antimelanoma IgG and 0.45 with the polyclonal IgG.

Clinical improvement after ventricular aneurysm repair: prediction by angiographic and hemodynamic variables.

Surgical repair of a left ventricular aneurysm is associated with significant perioperative mortality and substantial mortality in the first 2 years after operation. In a retrospective review of 42 patients undergoing repair of an anteroapical aneurysm, two cardiac catheterization variables were identified that predicted a good surgical outcome, defined as perioperative survival and improved functional status. Specifically, patients with an ejection fraction of the contractile section (nonaneurysmal) of the left ventricle of 35% or greater and a left ventricular end-diastolic pressure of 25 mm Hg or less had a low perioperative mortality rate (6.5%), experienced no late mortality and had sustained clinical improvement of at least one New York Heart Association functional class (93.5%). In contrast, patients with a contractile section ejection fraction of less than 35% or a left ventricular end-diastolic pressure greater than 25 mm Hg had a higher perioperative mortality rate (27.3%), experienced a substantial late mortality rate (27.3%) or had no significant functional class improvement (9%); only 36.4% had sustained clinical improvement. This study suggests that the postoperative results of left ventricular aneurysm repair are dependent on the hemodynamic status of the nonresected left ventricle.

Mechanisms of nondisjunction in human spermatogenesis.

A reduction in recombination in the pseudoautosomal region is associated with an increased frequency of aneuploid 24,XY human sperm. Similarly, individuals with paternally derived Klinefelter syndrome (47,XXY) also have a paucity of recombination in the chromosomes that have undergone nondisjunction. Meiotic studies using newly developed immunocytogenetic techniques have demonstrated errors of chromosome synapsis and significantly reduced recombination in infertile men with nonobstructive azoospermia. These men have an increased risk of aneuploidy in sperm that have been surgically removed from the testes. Thus, evidence is starting to accumulate that reduced recombination has a marked effect on the generation of aneuploid sperm.

Male infertility in reciprocal translocation carriers: the sex body affair.

Previous reports have linked chromosomal reorganization and spermatogenic failure. In this context, it has long been known that reciprocal translocation carriers are more likely to have anomalies in the meiotic process, including fertility failures. It has also been proposed that this fertility failure may be a consequence of an association between the translocated chromosomes and the sex body. In this work, we review different hypotheses explaining meiotic failure in these carriers, and propose a model that relates meiotic abnormalities with both sex body-translocation association and different checkpoints that are known to operate during meiosis.

Immunofluorescent synaptonemal complex analysis in azoospermic men.

The molecular cause of germ cell meiotic defects in azoospermic men is rarely known. During meiotic prophase I, a proteinaceous structure called the synaptonemal complex (SC) appears along the pairing axis of homologous chromosomes and meiotic recombination takes place. Newly-developed immunofluorescence techniques for SC proteins (SCP1 and SCP3) and for a DNA mismatch repair protein (MLH1) present in late recombination nodules allow simultaneous analysis of synapsis, and of meiotic recombination, during the first meiotic prophase in spermatocytes. This immunofluorescent SC analysis enables accurate meiotic prophase substaging and the identification of asynaptic pachytene spermatocytes. Spermatogenic defects were examined in azoospermic men using immunofluorescent SC and MLH1 analysis. Five males with obstructive azoospermia, 18 males with nonobstructive azoospermia and 11 control males with normal spermatogenesis were recruited for the study. In males with obstructive azoospermia, the fidelity of chromosome pairing (determined by the percentage of cells with gaps [discontinuities]/splits [unpaired chromosome regions] in the SCs, and nonexchange SCs [bivalents with 0 MLH1 foci]) was similar to those in normal males. The recombination frequencies (determined by the mean number of MLH1 foci per cell at the pachytene stage) were significantly reduced in obstructive azoospermia compared to that in controls. In men with nonobstructive azoospermia, a marked heterogeneity in spermatogenesis was found: 45% had a complete absence of meiotic cells; 5% had germ cells arrested at the zygotene stage of meiotic prophase; the rest had impaired fidelity of chromosome synapsis and significantly reduced recombination in pachytene. In addition, significantly more cells were in the leptotene and zygotene meiotic prophase stages in nonobstructive azoospermic patients, compared to controls. Defects in chromosome pairing and decreased recombination during meiotic prophase may have led to spermatogenesis arrest and contributed in part to this unexplained infertility.

True and false aneurysms of the left ventricle following myocardial infarction.

Anterolateral myocardial infarction resulted in the formation of both true and false aneurysms in a 75 year old man in whom severe congestive heart failure subsequently developed as the false aneurysm became progressively larger. Left ventriculography detected and quantified both aneurysms, and demonstrated reasonable function of the remaining volume-overloaded left ventricle. Resection of both aneurysms was accomplished with marked relief of symptoms. The literature on false aneurysm is reviewed, and the dilemma posed by the need to recognize false aneurysms before they become symptomatic or rupture is discussed.

The effect of cold storage on recombination frequencies in human male testicular cells.

Meiotic recombination is essential for the segregation of homologous chromosomes and formation of normal haploid gametes. Decreased recombination is associated with the production of aneuploid sperm in humans. MLH1, a DNA mismatch repair protein, was recently found to mark the sites of recombination in humans. Newly developed immunofluorescence techniques to identify MLH1 foci on synaptonemal complexes (SCs) in pachytene cells from testicular tissue have opened up a new avenue of research on meiotic recombination. Future studies on normal and abnormal recombination in early meiosis will further research in human reproduction and genetics. However, the availability of testicular material will always be a major limiting factor in this kind of study. In order to obtain an adequate number of samples and samples of particular research interest, it is often of benefit to obtain samples from distant regions. Therefore, it is necessary to determine whether the quality of samples and accuracy of MLH1 frequencies change after transporting testicular samples from a distance. In the present study, we examined the recombination frequencies (numbers of MLH1 foci using immunofluorescence techniques) in 6 normal testicular samples. Each sample was split and analyzed in the fresh state and after storage on ice for two days, mimicking overnight courier air transport. The results showed no significant difference in the quality of the SC preparations or in the number of MLH1 foci between these two groups. These results demonstrate that testicular specimens may be shipped on ice without compromising data on chromosome pairing and recombination in early meiosis.

Gallium-SPECT in the detection of prosthetic valve endocarditis and aortic ring abscess.

A 52-yr-old man who had a bioprosthetic aortic valve developed Staphylococcus aureus bacteremia. Despite antibiotic therapy he had persistent pyrexia and developed new conduction system disturbances. Echocardiography did not demonstrate vegetations on the valve or an abscess, but gallium scintigraphy using SPECT clearly identified a focus of intense activity in the region of the aortic valve. The presence of valvular vegetations and a septal abscess was confirmed at autopsy. Gallium scintigraphy, using SPECT, provided a useful noninvasive method for the demonstration of endocarditis and the associated valve ring abscess.

Gallium-67-citrate and bone scintigraphy in disseminated North American blastomycosis.

We present a patient with North American blastomycosis involving lung and bone. Chest radiographs and CT scan showed a mass in the lung. Bone scintigraphy detected a photon-deficient area in the sternum and 67Ga SPECT showed uptake in the right upper lung and in the sternum. A diagnostic thoracotomy and needle biopsy from the sternal lesion revealed granulomatous infection due to Blastomyces dermatitidis. After 3 mo of antifungal therapy, the follow-up 67Ga study showed no evidence of the original lesions but demonstrated a new, asymptomatic, unsuspected lesion in the left infraspinatous muscle. This case illustrates that North American blastomycosis should be included in the differential diagnosis in cases of atypical pulmonary disease with bone involvement, even in geographic regions that are not considered endemic for this microorganism. Gallium-67 and bone scintigraphy may be useful in determining the extent of dissemination, in detecting occult lesions and in the follow-up of response to therapy.

Indium-111 chloride scintigraphy in adult osteomyelitis.

Osteomyelitis is a common clinical problem that may be difficult to diagnose. We compared the performance of indium-111-labeled white cells ([111In]WBC) to 111In chloride ([111In]Cl) in two groups of adult patients suspected to have osteomyelitis or septic arthritis. Using [111In] WBC, 52 scans were performed on 51 patients. Nineteen patients had osteomyelitis. The sensitivity was 84% and specificity 82%. Using [111In]Cl, 48 scans were performed on 47 patients. Twelve had osteomyelitis. Sensitivity was 91%, and specificity was 89%. In each group, one false-negative study occurred in vertebral osteomyelitis. Three false-negative studies using [111In]WBC were due to failure to distinguish between combined bone and soft-tissue infection and soft-tissue infection alone. False-positive studies in both groups were due to overlying soft-tissue infection or inflammatory arthritis. Chi-squared test showed no significant difference in performance between the two agents. Indium-111 chloride is easier to prepare and use than [111In]WBC, which requires a time-consuming labeling process.

Indium-111-white blood cell scintigraphy in Crohn's patients with fistulae and sinus tracts.

METHOD: Indium-111-white blood cell (111In-WBC) images of 17 Crohn's patients with fistulae and sinus tracts were reviewed and correlated with radiographic results (n = 16 patients) and surgery (n = 16 patients), to characterize the scintigraphic appearance of fistulization and to determine the role of 111In-WBC scintigraphy in this clinical setting. These were compared to 50 consecutive abnormal 111In-WBC studies obtained in Crohn's patients with suspected active disease but no known fistulae or sinus tracts. RESULTS: Scintigraphic findings which suggested the presence of fistulae were: (1) the presence of concomitant intestinal and extraintestinal lesions and (2) the absence of luminal activity on delayed images when early images detected bowel activity. The extraintestinal lesions were the drainage site of the fistula (n = 7) or an accompanying abscess (n = 6). Absence of luminal activity occurred in seven patients with fistulae and in two without fistulae; two patients had a proximal colostomy, two patients had bowel obstruction and five patients had fistulae to the skin (n = 3) or between the ileum and distal colon (n = 2). The distribution of active bowel disease as assessed scintigraphically was in complete agreement with surgery in 14 of 17 cases (82%) compared to 9 of 15 cases (60%) when correlating radiographic assessment with surgery. All surgically proven abscesses were detected on 111In-WBC images. CONCLUSION: These results indicate that 111In-WBC scintigraphy adds useful information to radiographic studies that is essential for appropriate management of Crohn's patients with fistulae and sinus tracts.

Assessment of painful late effects of lumbar spinal fusion with SPECT.

UNLABELLED: The authors reviewed planar, SPECT and other contemporaneous radiologic images of the spine and the medical records of 33 patients with back pain after lumbar fusion surgery in order to determine the value of SPECT in the assessment of painful late effects of spinal fusion surgery. METHODS: Twenty-one patients had lateral fusion, nine patients had posterior fusion only and three patients had anterior and posterior fusions. There were 24 patients who had surgery more than 4 yr ago (late group, mean 11.8 yr) and 9 patients who had surgery less than 4 yr ago (early group, mean 17.8 mo). RESULTS: The most common SPECT abnormality in patients in the late group were lesions in the vertebral bodies and apophyseal joints in the free motion segments adjacent to the fused segments (62.5% of patients). Such lesions occurred in 46% of patients after lateral fusion, in 87.5% of patients after posterior fusion and in 67% of patients after posterior and anterior fusions. No SPECT abnormalities were detected in the fused segments in patients in the late group with solid lateral fusion but were detected in three patients with solid posterior fusion. These results correlate with biomechanical studies that have shown posterior fusion to produce the largest amount and lateral fusion to produce the least amount of stress in the free segments adjacent to the fusion. Lateral fusion was found to have a more stabilizing effect than posterior fusion. CONCLUSION: In addition to the already established value of SPECT in detecting painful pseudoarthrosis, our results indicate that SPECT is of value in the assessment of painful late effects of fusion.