Problems in evaluating male fertility: valuable factors in evaluating male fertility and normal values of seminal parameters

To determine the valuable factor for evaluating male fertility, a comparative study was done as to various seminal parameters between fertile and infertile groups. The fertile group consists of 57 proven fertile males and the infertile group consists of randomly chosen 67 infertile patients. Seminal parameters assessed were sperm concentration, motility, mean velocity, total sperm output, total motile sperm output, sperm morphology, acrosin activity and sperm penetration rate on zona-free hamster egg penetration assay (SPA). The infertile group was significantly different from the fertile group in every parameter except acrosin activity. However, the range of each parameter in the two groups overlapped each other. The diagnostic rate of each parameter, which is the percentage of an infertile male correctly diagnosed as infertile, was calculated by using 95% specificity threshold value of fertile males. The 95% specificity threshold values of sperm concentration, motility and % normal shaped sperm were 24.9 x 10(6)/ml, 34.9% and 55%, respectively, and they could be acceptable for the normal limit of seminal parameters. The diagnostic rate was highest in penetration rate (72.4%). In other words, penetration rate is the most valuable factor in various parameters for making a distinction between fertile and infertile males. Sperm motility and mean velocity showed the next highest diagnostic rate. On the other hand, sperm concentration showed a poor diagnostic rate (36.8%). In addition, there was no significant correlation between penetration rate and any other seminal parameters. These results suggest that the SPA will be an essential test for evaluating male fertility and penetration rate may be a marker of male fertility in the treatment of male infertility.     

Expression of human lymphocyte antigen (HLA)-DR on tumor cells in basal cell carcinoma

Immunohistologic studies of eight patients with basal cell carcinoma were undertaken using a series of monoclonal antibodies. In all of the patients, the majority of dermal infiltrates reacted with OKT3 and OKIa1 (HLA-DR), with a slight predominance of OKT4+ helper/inducer T cells (the mean OKT4/OKT8 ratio was 1.8). Both OKT4+ and OKT8+ cells were seen infiltrating the tumor masses. In addition, in five cases, human lymphocyte antigen (HLA)-DR was demonstrated on some tumor cells close to a vast number of HLA-DR+ infiltrates surrounding the carcinoma, but not on epidermal keratinocytes and tumor cells devoid of the HLA-DR+ infiltrates. A considerable number of OKT6+ dendritic cells were also observed surrounding the carcinoma. Staining with OKB7 and OKM1 revealed negligible reactive cells, and virtually none of the dermal infiltrates reacted with Leu-7 (HNK-1). These findings suggest that in addition to varied immunologically competent cells, expression of HLA-DR antigen on tumor cells may participate in a cellular immune reaction, a defense mechanism against tumor cell proliferation in basal cell carcinoma.     

Super high resolution for single molecule-sequence-based typing of classical HLA loci at the 8-digit level using next generation sequencers

Current human leukocyte antigen (HLA) DNA typing methods such as the sequence-based typing (SBT) and sequence-specific oligonucleotide (SSO) methods generally yield ambiguous typing results because of oligonucleotide probe design limitations or phase ambiguity for HLA allele assignment. Here we describe the development and application of the super high-resolution single-molecule sequence-based typing (SS-SBT) of HLA loci at the 8-digit level using next generation sequencing (NGS). NGS which can determine an HLA allele sequence derived from a single DNA molecule is expected to solve the phase ambiguity problem. Eight classical HLA loci-specific polymerase chain reaction (PCR) primers were designed to amplify the entire gene sequences from the enhancer-promoter region to the 3' untranslated region. Phase ambiguities of HLA-A, -B, -C, -DRB1 and -DQB1 were completely resolved and unequivocally assigned without ambiguity to single HLA alleles. Therefore, the SS-SBT method described here is a superior and effective HLA DNA typing method to efficiently detect new HLA alleles and null alleles without ambiguity.     

Electrophysiological studies on the cerebellocerebral projections in monkeys

1. Responses evoked by stimulation of the cerebellar and thalamic nuclei were recorded by microelectrodes introduced at various depths in the cerebral cortex of monkeys (Macaca mulatta) under light Nembutal anaesthesia. 2. Stimulation of the medial (fastigial) cerebellar nucleus produced, at a latency of 4-5 msec, deep thalamo-cortical (T-C) responses (surface positive-deep negative potentials) mainly in the medial part of the precentral gyrus (area 4, "motor area for hindlimb") and in the superior parietal gyrus (area 5) on both contralateral and ipsilateral sides to the nucleus stimulated. 3. Stimulation of the lateral (dentate) cerebellar nucleus elicited, at a latency of about 3 msec, superficial T-C responses (surface negative-deep positive potentials) predominately in the lateral part of the precentral gyrus (area 4, "motor area for forelimb and face") and in the rostromedial part of the gyrus (area 6, premotor area) on the contralateral side. 4. Stimulation of the interpositus cerebellar nucleus set up superficial T-C responses chiefly in the motor area between those influenced by the medial and the lateral cerebellar nucleus stimulation and also in the premotor area on the contralateral side. 5. The respective areas responsive to the medial, interpositus and lateral nucleus stimulation overlapped considerably each other in the motor cortex. 6. Comparison of the responses in the cortex induced by stimulation of the cerebellar and thalamic nuclei indicated different relay portions in and around the VA-VL region of the thalamus for the superficial and the deep T-C responses respectively. 7. Functional implications of the results were discussed in referring to the cerebellocerebral projections in cats.     

Evaluation of Combinations of 4′-Ethynyl-2-Fluoro-2′-Deoxyadenosine with Clinically Used Antiretroviral Drugs

Drug combination studies of 4′-ethynyl-2-fluoro-2′-deoxyadenosine (EFdA) with FDA-approved drugs were evaluated by two different methods, MacSynergy II and CalcuSyn. Most of the combinations, including the combination of the two adenosine analogs EFdA and tenofovir, were essentially additive, without substantial antagonism or synergism. The combination of EFdA and rilpivirine showed apparent synergism. These studies provide information that may be useful for the design of EFdA combination regimens for initial and salvage therapy assessment.     

Stem Cell Therapies in Patients with Single Ventricle Physiology

Single ventricle physiology, especially hypoplastic left heart syndrome, is one of the most high-risk lesions in children with congenital heart disease, and the ensuing heart failure remains as a major problem related to adverse outcomes in these patients. The field of stem cell therapy for heart failure has shown striking advances during the past 10 years, and many clinical trials using stem cell technologies have been conducted in adults, which suggest that stem cell therapy is associated with long-term improvement in cardiac function. Cardiac progenitor cells have recently been discovered, and their strong regenerative ability has been demonstrated in several studies. Although no large clinical trials have been performed in the field of congenital heart disease, recent investigations indicate that stem cell therapy may hold great potential to treat children with cardiac defects.     

Impact of age on the prognostic value of number of lymph nodes retrieved in patients with stage II colorectal cancer

Purpose

A small number of lymph nodes retrieved (NLNR) is a known risk factor in stage II colorectal cancer. NLNR is influenced by age, but little is known about whether the impact of small NLNR on survival differs with age. This retrospective study sought to determine such impact in elderly patients with stage II colorectal cancer.

Methods

We reviewed data for 2100 patients with stage II colorectal cancer who underwent surgery without adjuvant chemotherapy between January 1997 and December 2003. The optimal cutoff value of NLNR for survival was determined, and the impact of small NLNR on survival was analyzed. The association between age and NLNR was evaluated. The relation between age and risk of small NLNR with respect to survival was then assessed to determine the impact of small NLNR on elderly patients’ survival.

Results

The optimal cutoff value of NLNR was determined as 6. The small NLNR group (SNG) showed significantly worse prognosis than the large NLNR group (LNG) (p?<?0.001). Age, surgical method, and scope of lymph node dissection were significantly associated with NLNR. A potential interaction was noted between age and risk of small NLNR in relation to relapse-free survival (RFS). Five-year RFS was significantly worse in SNG than in LNG for elderly patients (41.7 and 76.4 %, respectively; p?<?0.001) but not for non-elderly patients (75.9 and 84.6 %, respectively; p?=?0.083).

Conclusions

NLNR <6 was identified to be an important prognostic factor for elderly patients with stage II colorectal cancer.

     

Analysis of c-myc DNA amplification in non-small cell lung carcinoma in comparison with small cell lung carcinoma using polymerase chain reaction

Previous studies of c-myc DNA amplification in lung cancer have focused primarily on analysis of small cell carcinoma or its tumor cell lines. There are few data about c-myc DNA amplification in histological types of lung cancer other than small cell carcinoma. Therefore the present study was conducted to investigate c-myc oncogene amplification in non-small cell lung carcinoma. We studied 46 lung tumor specimens for c-myc DNA amplification (15 adenocarcinomas, 15 squamous cell carcinomas, 6 large cell carcinomas, and 10 small cell carcinomas). Polymerase chain reaction, digoxigenin DNA labeling, and elecrophoresis were utilized to investigate the c-myc copy number in the lung tumor specimens. The c-myc copy number of non-small cell carcinoma ranged from 1.5 to more than 20.0 in adenocarcinoma and squamous cell carcinoma, and from 6.0 to 12.0 in large cell carcinoma. That of small cell carcinoma ranged from 1.8 to 12.0. The c-myc copy number of non-small cell carcinoma was significantly higher that than of small cell carcinoma (Wilcoxon rank sum test, Z=2.06, P=0.040). However, the differences in c-myc copy number among these four histological types were not statistically significant. Amplification of c-myc (more than 4 copies) was observed not only in small cell carcinoma but also in non-small cell carcinoma at similarly high frequency (12/15 in adenocarcinoma and squamous cell carcinoma, 6/6 in large cell carcinoma, and 9/10 in small cell carcinoma). Received: 17 October 2000 / Accepted: 7 May 2001