bluuebunny/crossref_metadata_2025 huggingface dataset:
Apical extrusion of debris and irrigants using two hand and three engine‐driven instrumentation techniques
Authors: Ferraz C. C. R. , Gomes N. V. , Gomes B. P. F. A. , Zaia A. A. , Teixeira F. B. , Souza‐Filho F. J.
Published: March 2003

Aim

To evaluate the weight of debris and irrigant volume extruded apically from extracted teeth in vitro after endodontic instrumentation using the balanced force technique, a hybrid hand instrumentation technique, and three engine‐driven techniques utilizing nickel–titanium instruments (ProFile .04, Quantec 2000 and Pow‐R).

Methodology

Five groups of 20 extracted human teeth with single canals were instrumented using one or other of five techniques: balanced force, hybrid, Quantec 2000, ProFile .04, or Pow‐R. Debris extruded from the apical foramen during instrumentation were collected into preweighed 1.5 mL tubes. Following instrumentation, the volume of extruded irrigant fluid was determined by visual comparison to control centrifuge tubes filled with 0.25 mL increments of distilled water. The weight of dry extruded dentine debris was also established.

Results

Overall, the engine‐driven techniques extruded less debris than the manual ones. However, there was no statistical difference between the balanced force technique and the engine‐driven methods. The volume of irrigant extruded through the apex was directly associated with the weight of extruded debris, except within the ProFile group. The hybrid technique was associated with the greatest extrusion of both debris and irrigant.

Conclusions

Overall, the engine‐driven nickel–titanium systems were associated with less apical extrusion.
Detection of Slackia exigua, Mogibacterium timidum and Eubacterium saphenum from pulpal and periradicular samples using the Polymerase Chain Reaction (PCR) method
Authors: Hashimura T. , Sato M. , Hoshino E.
Published: March 2003

Aim

The purpose of this study was to detect Slackia exigua from root canal samples using a sensitive PCR amplification method. Mogibacterium timidum and Eubacterium saphenum were also included because of their culture‐difficult properties.

Methodology

The species‐specific PCR primers were prepared according to 16S rDNA sequence analysis data, and confirmed to be effective for PCR amplification as species‐specific, respectively. A total of 36 clinical samples were obtained during the first visit of root canal treatment.

Results

The sensitivity of detection was a minimum of 10 organisms for S. exigua and five organisms for M. timidum and E. saphenum, respectively. In seven cases of pulpitis, S exigua was detected in two cases (29%), and M. timidum in two cases (29%), but E. saphenum was not detected. In 17 cases of root canal treatment, S.exigua was detected in seven cases (41%), M. timidum in 12 cases (71%) and E. saphenum in four cases (24%). In 12 cases of root canal retreatment, S. exigua was detected in three cases (25%), M. timidum in three cases (25%) and E. saphenum in two cases (17%).

Conclusions

S. exigua, M. timidum and E. saphenum were present in root canal systems, and may be associated with pulpal and periradicular pathosis.
The quality of root fillings remaining in mandibular incisors after root‐end cavity preparation
Authors: Wu M‐K. , De Schwartz F. B. C. , Van Der Sluis L. W. M. , Wesselink P. R.
Published: March 2003

Aim

The aim of this study was to determine the quality of root fillings remaining in mandibular incisors after root‐end resection and root‐end cavity preparation.

Methodology

Roots of 40 mandibular incisors, 12 mm in length, were divided into two groups and instrumented using a balanced force technique and obturated by vertically compacted warm gutta‐percha. In one group sealer was used when back‐filling the root canal with the Obtura System. The other group was also back‐filled with the Obtura System but without sealer. The apical 4 mm of each root filling was removed by root‐end resection and root‐end cavity preparation. Fluid transport along the 7 mm of remaining root filling was measured. After the leakage test, horizontal sections were cut at a level 3 mm from the apical end of the remaining root filling. The cross sectional area of the canal and gutta‐percha was measured using an image analysis program. The quality of the remaining root fillings was also evaluated by calculating the percentage of gutta‐percha filled canal area.

Results

In total, 27 (67%) out of the 40 roots demonstrated leakage. No significant difference was found between the two groups (P = 0.265). In the cross‐sections the bucco‐lingual canal diameter was up to 2.6 mm long (mean 1.6 mm). The average percentage of gutta‐percha filled canal area was 85.6%. Warm gutta‐percha could not obturate areas where debris remained after instrumentation.

Conclusions

The quality of the root fillings in mandibular incisors was poor, probably because the unprepared recesses of oval canals were not clean.
A case of unusual anatomy: a mandibular second premolar with four canals
Authors: Rhodes J. S.
Published: March 2003

Aim

The aim of this case report is to describe conventional root canal treatment on a mandibular second premolar with four canals.

The pulp of a mandibular second premolar had become irreversibly inflamed, probably as a result of bacterial microleakage from a carious lesion. The general dental practitioner had initiated root canal treatment, but due to procedural difficulties referred the case. Conventional root canal treatment was then performed with magnification. Preparation was undertaken in a crown‐down manner using balanced force hand instrumentation with flexible K‐type files and files of Greater Taper. The root canals were obturated with vertically condensed gutta‐percha technique.

Key learning points

Good illumination and magnification are vital during root canal treatment.

Crown‐down preparation using the balanced force technique with flexible files makes instrumentation easier and prevents many of the procedural errors that can occur with filing techniques.

Greater Taper instruments made of nickel–titanium are sufficiently flexible to be used in complex curved canals and obviate the need for step‐back flaring of the apical preparation.

Vertical compaction of warm gutta‐percha may simplify the obturation of complex root canal systems.

When the technical difficulty of a procedure exceeds the expertise of the practitioner, specialist referral may be required.
Quantitative analysis of substance P, neurokinin A and calcitonin gene‐related peptide in pulp tissue from painful and healthy human teeth
Authors: Awawdeh L. , Lundy F. T. , Shaw C. , Lamey P‐J. , Linden G. J. , Kennedy J. G.
Published: March 2003

Aim The purpose of this study was to investigate the levels of substance P (SP), neurokinin A (NKA) and calcitonin gene‐related peptide (CGRP) in painful and healthy human dental pulps.

Methodology Forty‐six samples of pulp tissue were collected from extracted or endodontically treated painful teeth and 20 from clinically healthy teeth extracted for orthodontic reasons. All pulp samples were boiled in 0.5 m acetic acid for 10 min, centrifuged and the supernatant collected. SP, NKA and CGRP levels were measured using radioimmunoassay.

Results Substance P and CGRP were present in all samples and NKA was detected in 96% of the pulps. CGRP was present in much higher concentrations than SP and NKA in both painful and non‐painful teeth. The painful teeth had significantly higher concentrations of SP (P = 0.02), NKA (P < 0.001) and CGRP (P = 0.03) than non‐painful teeth. The concentration of CGRP was significantly higher in the pulps of smokers compared with non‐smokers (P = 0.02).

Conclusions Elevated levels of these neuropeptides in pulps from painful teeth indicate that they may play an important role in the process of pulpal inflammation and pain. Further investigation of the association between these neuropeptides and pulpal status may help to improve our understanding of pulpal inflammation and dental pain.
Interleukin‐1β and tumour necrosis factor‐α levels in periapical exudates
Authors: Ataoğlu T. , Üngör M. , Serpek B. , Haliloğlu S. , Ataoğlu H. , Ari H.
Published: March 2003

Aim The aim of this study was to determine IL‐1β and TNF‐α levels in periapical exudates and to evaluate their relationship with clinical and radiological findings.

Methodology Periapical exudates were collected from root canals of 35 single‐rooted teeth using absorbent paper points. IL‐1β and TNF‐α levels were determined by enzyme‐linked immunosorbent assays. The samples were divided into two groups according to the presence or absence of clinical signs including swelling and/or fistula, pain on palpation and/or percussion, and pus discharge from canals. Periapical exudate samples were divided into two groups according to size of periapical radiolucent area.

Results The mean concentration of IL‐1β (72.79 ng mL−1) in periapical exudates was approximately 12‐fold higher than TNF‐α (6.17 ng mL−1). There was no significant correlation between these cytokines (P > 0.05). IL‐1β levels in canals with larger radiolucent areas (long axis ≥1 cm) were significantly higher than those with small areas (P < 0.05). There was a tendency toward higher levels of IL‐1β in groups with clinical signs, but the differences were not significant (P > 0.05).

Conclusions Periapical exudate levels of both cytokines failed to reflect periapical disease state.
Influence of age, hormones and germ cells on glutathione S‐transferase activity in cultured Sertoli cells
Authors: CASTELLÓN
Published: March 2003

Glutathione S‐transferase (GSH‐S‐T) activity was measured, using 1‐Cl‐2,4‐dinitrobenzene as substrate, in Sertoli cell cultures obtained from rats aged 10, 18, and 26 days. The GSH‐S‐T activity showed a significant increase with age of the Sertoli cell donor. When cultures were treated with hypotonic solution, in order to eliminate residual contaminating germ cells, the age dependent increase in enzyme activity was less pronounced. FSH, but not testosterone, increased enzyme activity in all cultures. Addition of freshly isolated germ cells (mainly pachytene spermatocytes) to hypotonic‐treated Sertoli cell monolayers enhanced GSH‐S‐T activity at all ages.

It is concluded that GSH‐S‐T activity can be measured in cultured Sertoli cells during the period of onset of spermatogenesis (10–26 days). This enzyme activity is dependent on age of the Sertoli cell donor and is influenced by FSH and germ cells. Since GSH‐S‐Ts are actively engaged in cell detoxificative functions through conjugation of xenobiotics with glutathione, the present findings suggest that this enzyme may have a relevant protective role during the critical period when spermatogenesis is being established.
Effect of freeze–thawing procedure on chromatin stability, morphological alteration and membrane integrity of human spermatozoa in fertile and subfertile men
Authors: HAMMADEH  , ASKARI  , GEORG  , ROSENBAUM  , SCHMIDT
Published: March 2003

Cryopreservation is known to impair sperm motility and decrease the fertilization rate by detrimental effects on acrosomal structure and acrosin activity. However, the consequences of cryopreservation on the integrity of the sperm nucleus, chromatin stability and centrosome are less clear. The present study was designed to determine the effect of the freeze–thawing procedure on chromatin condensation (aniline blue staining) and the morphology (strict criteria) and membrane integrity of human spermatozoa. The structural and functional characteristics of the sperm plasma membrane were measured by the eosin‐test and hypo‐osmotic swelling test which were done separately. Sperm cryopreservation was performed on semen samples from two groups of men classified as fertile (n = 20) and subfertile (n = 72), based on their reproductive history and semen analysis according to WHO guidelines. The mean percentage of condensed chromatin, morphologically normal spermatozoa and membrane integrity in all semen samples investigated (n = 92) decreased significantly (p = 0.0001) after freeze–thawing, in comparison to the value observed prior to freezing. By comparing the semen samples between fertile and subfertile patients, significantly (p = 0.0009) greater damage was demonstrated in the subfertile than in the fertile group. Furthermore, no significant difference was observed between the two groups with regard to the morphological alteration and structural as well as functional damage of the sperm membrane. In conclusion, the freeze–thawing procedure significantly affects chromatin structure and sperm morphology, especially in the head and the tail regions, and this may explain the lower fertilization rate and IVF/ICSI outcome when frozen–thawed spermatozoa are used. In addition, this study demonstrates that chromatin condensation is a sensitive parameter for the evaluation of cryodamage of semen samples from fertile and subfertile patients, though subfertile patients with very poor semen characteristics have yet to be studied. It is therefore recommended that chromatin condensation be used as an additional parameter for the assessment of sperm quality after freeze–thawing.
GH/IGF‐I axis in azoospermia in primary and secondary hypogonadism: a study before and during replacement therapy
Authors: CARANI  , MANTOVANI  , PROCOPIO  , RIO G. DEL , ROSSETTO  , GRANATA
Published: March 2003

The growth hormone/insulin‐like growth factor‐I (GH/IGF‐I) axis was studied in 15 azoospermic patients and in 10 control men. Eight patients were affected by hypergonadotrophic hypogonadism and 7 by hypogonadotrophic hypogonadism. All were studied before and during replacement therapy with testosterone and gonadotrophin, respectively, using the α2 adrenergic agonist, clonidine (clonidine test). The data demonstrate no differences in basal levels for IGF‐I and for the GH response to clonidine in azoospermic patients, affected by primary and secondary hypogonadism, before and during replacement therapy when compared with control fertile men. In contrast to some studies which describe a reduced GH response in azoospermia and oligozoospermia, we conclude that basal serum levels of IGF‐I and the GH response to clonidine are not impaired in azoospermic patients affected by primary hypogonadism before and after the restoration of normal androgenization, and in azoospermic patients affected by secondary hypogonadism, both before and after restoration of spermatogenesis.
Exposure to diesel exhaust affects the male reproductive system of mice
Authors: Yoshida Seiichi , Sagai Masaru , Oshio Shigeru , Umeda Takashi , Ihara Tomomi , Sugamata Masao , Sugawara Isamu , Takeda Ken
Published: March 2003

Several recent reports have suggested that sperm count and quality in normal men are declining. Various environmental chemical compounds may affect the male reproductive system. We propose here that diesel exhaust is an environmental pollutant with the potential to influence male reproductive function. Ultrastructural changes were observed in Leydig cells of mice exposed to diesel exhaust (0.3 mg diesel exhaust particles (DEP)/m3 through the airway, 12 h daily, up to 6 months) and reduction in LH receptor mRNA expression in Leydig cells was observed at a concentration of 1 mg DEP/m3. Daily sperm production per gram of testis dose‐dependently decreased with exposure to DE for 6 months; 29%, 36%, and 53% reductions were observed at 0.3, 1.0, and 3.0 mg DEP/m3, respectively. A no‐observed‐adverse‐effect level (NOAEL) was observed with approximately 30 μg DEP/m3, which is lower than the WHO‐recommended limit.
Comparative study of disomy and diploidy rates in spermatozoa of fertile and infertile men: a donor‐adapted protocol for multi‐colour fluorescence in situ hybridization (FISH)
Authors: Schultz Hendrik , Mennicke Kirsten , Schlieker Heiko , Al‐Hasani Safat , Bals‐Pratsch Monika , Diedrich Klaus , Schwinger Eberhard
Published: March 2003

Spermatozoa from seven healthy donors (two of whom had already fathered children) and five infertile patients taking part in the local programme of intracytoplasmic sperm‐injection (ICSI) were investigated for the disomy rates of chromosomes 13/21, 18, X and Y as well as for the diploidy rates. Two‐ and three‐colour fluorescence in situ hybridization (FISH) was applied after a donor‐adapted decondensation pre‐treatment: in a preliminary decondensation series the optimum fluorescence signals were individually determined by variation of the concentration of the decondensation reagents and the duration of incubation with these reagents. Strict scoring criteria were applied.

The average disomy rates ranged from 0.10% (chromosomes 13/21) to 0.44% (disomy XY) in the infertile donors and from 0.07% (disomy XX) to 0.36% (disomy XY) in the controls. The average diploidy rates were 0.22% and 0.20% for the infertile donors and the controls respectively. There was no statistically significant difference between the two groups with respect to the disomy and diploidy rates. Within the two groups there were inter‐individual differences which were partly statistically significant, indicating considerable inter‐donor variation of the aneuploidy rates.
Prediction of unexpectedly poor fertilization and pregnancy outcome using the strict criteria for sperm morphology before and after sperm separation in IVF–ET
Authors: Obara H. , Shibahara H. , Tsunoda H. , Taneichi A. , Fujiwara H. , Takamizawa S. , Idei S. , Sato I.
Published: March 2003

This study was performed to investigate if unexpectedly poor fertilization and in‐vitro fertilization (IVF) outcome could be predicted using sperm morphology as diagnosed by the strict criteria. Sperm morphology was assessed in 137 IVF–ET cycles with at least three oocytes collected. The lowest amount of normal forms was 5% in 137 samples, indicating there were no patients belonging to ‘poor prognosis’ (<5% normal forms). Treatment using intracytoplasmic sperm injection (ICSI) was also excluded. Before sperm separation by the swim‐up method, sperm morphology demonstrated a significant correlation with the fertilization rate (p < 0.0001). The fertilization rate (80.5%) in 110 ‘normal’ samples (>14% normal forms) was significantly higher (p < 0.01) than that (55.4%) in 27 samples with ‘good prognosis’ (those with 5–14% normal forms). No embryo was available for transfer (ET) in 4 (3.6%) of 110 ‘normal’ cycles and in 3 (11.1%) of 27 ‘good prognosis’ cycles (not significant). Fresh ET was intentionally cancelled to avoid severe ovarian hyperstimulation syndrome (OHSS) in six of 110 ‘normal’ cycles as well as in one of 27 ‘good prognosis’ cycles. The pregnancy rate per ET was 31.0% (31/100) in the former group, while it was 26.1% (6/23) in the latter group. There was no difference between the two groups. In the post swim‐up evaluation of sperm characteristics, morphology was significantly correlated with the fertilization rate in IVF–ET (p < 0.05) while other sperm parameters were not. When the cut off level for the post swim‐up sperm morphology was set at 25%, there was a significant difference in the fertilization rates between patients (78.6%) with post‐swim‐up >25% and those (55.0%) with post‐swim‐up ≤25% (p < 0.01). Taken together, a relative indication for ICSI using sperm morphology before and after swim‐up was established. Category A includes ≤14% normal forms in the ejaculate and post‐swim‐up ≤25%, while Category B includes ≤14% in the ejaculate and post‐swim‐up >25%. There was a significant difference in the fertlization rates between patients (47.2%) in Category A and those (60.2%) in Category B (p < 0.05). The clinical pregnancy rate was 11.1% for patients in Category A compared with 35.7% for patients in Category B. However, there was no significant difference between the two categories. These results indicate that the strict criteria provide a reliable estimation of the fertilizing ability of human spermatozoa. ICSI might be considered in Category A patients to avoid poor fertilization and pregnancy outcome.
Assay of testicular angiotensin‐converting enzyme activity in human spermatozoa
Authors: Kamata Makiko , Hu Jianguo , Shibahara Hiroaki , Nakagawa Hachiro
Published: March 2003

The testicular isozyme of angiotensin‐converting enzyme (ACE) is associated with male fertility. Spermatozoa from mice lacking ACE showed defects in transport within the oviducts and in binding to zonae pellucidae although the animals had normal sperm count, morphology and motility. In fact, unexplained infertility is difficult to be predicted by conventional parameters such as sperm count. We measured membrane testicular ACE activity in a sperm suspension in PBS and total testis ACE activity in spermatozoa by solubilization with Triton X‐100. Total testis ACE activity and membrane testis ACE activity of the same subject were compared in 12 control subjects. We demonstrated that testicular ACE is stable in spermatozoa and the assay of testicular ACE activity is possible. Total testicular ACE activity was approximately twice the membrane testicular ACE activity in all of the subjects tested. The assay of testicular ACE activity in human spermatozoa could be a new method for the assessment of sperm function.
Effect of human sperm exposure to progesterone on sperm–oocyte fusion and sperm–zona pellucida binding under various experimental conditions
Authors: Francavilla F. , Romano R. , Santucci R. , Macerola B. , Ruvolo G. , Francavilla S.
Published: March 2003

In this study, the effect of human sperm exposure to progesterone on sperm/oocyte fusion, using the hamster egg penetration test, and on sperm/zona pellucida (ZP) binding, using the hemizona assay, was investigated under various experimental conditions.

A brief exposure of human spermatozoa to progesterone exerted a stimulatory effect on sperm/oocyte fusion which was dose‐dependent, capacitation‐dependent, influenced by the source of serum albumin in capacitating medium, and was higher than that produced by the exposure to progesterone from the onset of capacitation. The exposure of capacitated spermatozoa to progesterone during 20 min‐spermatozoa/ZP‐coincubation produced an enhancement of ZP‐binding, which was not significantly influenced by the source of serum albumin in capacitating medium. A significantly lower ZP‐binding was exhibited by spermatozoa exposed to progesterone from the beginning of capacitation. These results indicate that progesterone exerts a stimulatory effect on human sperm's fertilizing ability, which occurs mainly in post‐capacitation events directly involved in sperm/oocyte fusion and in ZP‐binding. Conditions optimizing these effects are provided. They should be taken into account in the standardization of experimental and clinical studies designed to evaluate the response of human spermatozoa to progesterone.
Effects of dihydrotestosterone on cardiac inward rectifier K+ current
Authors: Carnes Cynthia A. , Dech Spencer J.
Published: March 2003

There are well‐described sexually dimorphic differences both in the electrocardiogram and in the propensity to develop drug‐induced arrhythmias. The QT interval and the risk of ventricular proarrhythmia are reduced in males compared with females. Inward rectifier potassium current (IK1) is a primary determinant of the ventricular resting membrane potential, and an important contributor to myocardial excitability.Methods and results: Using the whole‐cell patch‐clamp technique, we evaluated the effects of dihydrotestosterone (DHT) on IK1 in ventricular myocytes from castrated rabbits that were treated with either replacement DHT or vehicle‐control for 3 weeks. Compared with the DHT‐treated group, myocytes from the control animals had a significant reduction in inward IK1 conductance (p < 0.005) and rectification ratio (RR) (p < 0.04) with no significant change in peak outward current. Acute DHT superfusion of the myocytes increased inward IK1 conductance from baseline (p < 0.05) and increased the RR (p < 0.05). Testosterone has been reported to increase intracellular ornithine decarboxylase activity in ventricular tissue, which would increase intracellular polyamines, known modulators of IK1 rectification. We found that inclusion of the intracellular polyamines spermidine and putrescine in the pipette solution caused a decrease in inward IK1, accompanied by an increase in peak outward current and a reduction in the RR. Conclusion: In summary, DHT modulates IK1 in a chronic, as well as, an acute fashion. These effects are not because of altered intracellular polyamines. DHT may modulate myocardial excitability through effects on IK1.
Management of premature ejaculation – a comparison of treatment outcome in patients with and without erectile dysfunction
Authors: Chia Sing joo
Published: March 2003

This study evaluated the problem of premature ejaculation (PE) in patients treated for erectile dysfunction. The aim was to compare the efficacy of selective serotonin reuptake inhibitors (SSRIs) in the management of primary PE and PE associated with sildenefil treatment. Eighty‐seven patients with PE seen over a period of 17 months were recruited into this prospective study. They were categorized into two groups: primary PE (GPI) and PE in sildenefil‐treated patients (GPII). All patients recruited into GPII had erectile dysfunction (ED) that was successfully treated with sildenefil citrate for at least a year. Both groups of patients were given sertraline 50 mg 4 h before expected time of sex. The minimum follow‐up was 6 months. The ejaculation latency before and after treatment of the two groups were compared. The sexual satisfaction scores of the patients in the two groups were also sought and analysed. Twenty‐eight percent of patients with ED who were successfully treated with sildenefil developed PE. Subjects in group GPI were younger and have less comorbid factors than those in group GPII. There was no significant difference in the mean ejaculation latency for both groups (46 vs. 34.6 sec for GPI and GPII, respectively). However, there was highly significant difference in the ejaculation latency between the two groups after treatment with sertraline for 6 months (247.2 vs. 111.6 sec for GPI and GPII, respectively). There was also significant difference in the sexual satisfaction score for group GPI post‐treatment, but not for GPII. No significant side‐effect of sertraline was reported from patients in both groups. Successful treatment of ED could not assure sexual satisfaction. At least a quarter of sildenefil treated ED patients might develop PE which would continue to frustrate these patients sexually. While selective serotonin re‐uptake inhibitors (SSRIs) was effective in the management of primary PE, they were not as effective in patients with sildenefil corrected ED.
Penile vibratory stimulation and electroejaculation in the treatment of ejaculatory dysfunction1
Authors: SØNKSEN JENS , OHL DANA A.
Published: March 2003

The purpose of this review is to present the current understanding of penile vibratory stimulation (PVS) and electroejaculation (EEJ) procedures and its clinical use in men with ejaculatory dysfunction. Unfortunately, the record of treating such individuals has been quite poor, but within recent years development and refinement of PVS and EEJ in men with spinal cord injury (SCI) has significantly enhanced the prospects for treatment of ejaculatory dysfunction. The majority of spinal cord injured men are not able to produce antegrade ejaculation by masturbation or sexual stimulation. However, approximately 80% of all spinal cord injured men with an intact ejaculatory reflex arc (above T10) can obtain antegrade ejaculation with PVS. Electroejaculation may be successful in obtaining ejaculate from men with all types of SCI, including men who do not have major components of the ejaculatory reflex arc. Because vibratory stimulation is very simple in use, non‐invasive, it does not require anaesthesia and is preferred by the patients when compared with EEJ, PVS is recommended to be the first choice of treatment in spinal cord injured men. Furthermore, EEJ has been successfully used to induce ejaculation in men with multiple sclerosis and diabetic neuropathy. Any other conditions which affect the ejaculatory mechanism of the central and/or peripheral nervous system including surgical nerve injury may be treated successfully with EEJ. Finally, for sperm retrieval and sperm cryopreservation before intensive anticancer therapy in pubertal boys, PVS and EEJ have been successfully performed in patients who failed to obtain ejaculation by masturbation. Nearly all data concerning semen characteristics in men with ejaculatory dysfuntion originate from spinal cord injured men. Semen analyses demonstrate low sperm motility rates in the majority of spinal cord injured men. The data give evidence of a decline in spermatogenesis and motility of ejaculated spermatozoa shortly after (few weeks) an acute SCI. Furthermore, it is suggested that some factors in the seminal plasma and/or disordered storage of spermatozoa in the seminal vesicles are mainly responsible for the impaired semen profiles in men with chronic SCI. Home insemination with semen obtained by penile vibratory and introduced intravaginally in order to achieve successful pregnancies may be an option for some spinal cord injured men and their partners. The majority of men will further enhance their fertility potential when using either penile vibratory or EEJ combined with assisted reproduction techniques such as intrauterine insemination or in‐vitro fertilization with or without intracytoplasmic sperm injection.
Expression and regulation of the prointerleukin‐1α processing enzymes calpain I and II in the rat testis
Authors: Sultana Taranum , Wahab‐Wahlgren Aida , Assmus Monique , Parvinen Martti , Weber Günther , Söder Olle
Published: March 2003

Interleukin‐1α (IL‐1α) is constitutively expressed in an age‐ and stage‐dependent manner by rat Sertoli cells. However, the mechanism of regulation of IL‐1α is unclear in testis. We studied this regulation at the level of the enzyme calpain, a potential regulator that cleaves 32 kDa proIL‐1α to produce mature 17 kDa IL‐1α. Both calpain I and II were found to cleave recombinant rat testis 32proIL‐1α in vitro. A temporary age‐related increase in messenger RNA (mRNA) levels of calpain I was found in testis of 20‐ and 25‐day‐old rats, coinciding with important events of spermatogenesis and a gradual increase in IL‐1α, while calpain II expression was constant. In response to lipopolysaccharide (LPS), calpain I protein levels were down‐regulated in the seminiferous tubules, while calpain II was less affected. By contrast, the liver after LPS treatment showed up‐regulated calpain I and II immunoreactive protein and reverse transcriptase chain reaction (RT‐PCR) signal. Depleting Leydig cells by ethane 1,2‐dimethane sulphonate treatment resulted in down‐regulated calpain I mRNA and protein expression, whereas calpain II remained unchanged. In summary, there is a differential expression of calpain I and II under pathological conditions induced either by endotoxin stimuli or Leydig cell depletion, which may produce a differential effect on IL‐1α processing.
Absence of association of androgen receptor trinucleotide expansion and poor semen quality
Authors: Erasmuson Tanya , Sin Iris L. , Sin Frank Y. T.
Published: March 2003

This study investigated the relationship between variation in the polymorphic CAG trinucleotide repeat (TNR) region of the human androgen receptor (AR) gene and semen quality in a Caucasian sample population. These men were patients attending the New Zealand Centre for Reproductive Medicine in Christchurch. The AR TNR region was amplified by polymerase chain reaction and then DNA sequenced to determine exact numbers of CAG repeats for each sample. In addition, the samples were screened for microdeletions within the AZFc region of the Y‐chromosome. A total of 105 men with poor semen quality were compared with a group of 93 men with normal semen quality. Men with poor semen quality had similar CAG repeat number to men with normal semen quality (21.46 ± 0.30 vs. 20.99 ± 0.28, p = 0.126). Y‐chromosome microdeletions were only detected in men with suboptimal semen parameters (7.4%). However, the presence of a deletion was not related to CAG repeat number. The CAG repeat number in the men with normal semen quality in the present study is similar to the Australian and German samples, but lower than those reported for the Swedes, Dutch and Danes. These results are contrary to the hypothesis that higher CAG repeats are associated with infertility in men, but strongly suggest that different populations may show different numbers of CAG repeats in addition to racial variation reported in previous studies.
Low cytoplasmic pH causes fragmentation and dispersal of the Golgi apparatus in human hepatoma cells
Authors: Yoshida Toshimichi , Kamiya Tomoko , Imanaka‐Yoshida Kyoko , Sakakura Teruyo
Published: March 2003

The centrosomal localization of the Golgi apparatus in interphase cells is thought to be maintained by retrograde microtubule‐based motility. It is well established that, when intracellular pH is lowered, lysosomes and endosomes, also showing pericentrosomal localization, translocate towards the plus ends of microtubules within 15 min. In this study, we found that prolonged incubation in low pH medium (pH 6.6) with 20 mm Na acetate induced the fragmentation and dispersal of the Golgi apparatus in the human hepatoma cell line PLC/PRF/5. The fraction of Golgi‐dispersed cells increased in a time‐dependent manner, and reached over 60% after the 16‐h incubation. The cytoplasmic pH was dropped to approximately 7.10. Replacement with normal pH medium restored the structure and localization of the apparatus within 30 min. In the low pH condition, the microtubular network and endoplasmic reticulum appeared normal, and cytoplasmic dynein was still bound to the fragmented Golgi membranes. These findings suggest that low cytoplasmic pH suppresses the retrograde movement of the Golgi apparatus as well as that of lysosomes and endosomes.
Cyclophilins and their possible role in the stress response
Authors: Andreeva Larisa , Heads Richard , Green Colin J.
Published: March 2003

Cyclophilins are proteins which are remarkably conserved through evolution; moreover they have been found in every possible existing organism, which indicates their fundamental importance. Due to their enzymatic properties, multiplicity, cellular localization and role in protein folding they belong to the group of proteins termed molecular chaperones. All the proteins of the cyclophilin family possess enzymatic peptidyl‐prolyl isomerase activity (PPI‐ase), which is essential to protein folding in vivo. Recently PPI‐ase activity was suggested as playing a role in regulation of transcription and differentiation. However, not all cyclophilin functions are explained by PPI‐ase activity. For instance, one of the cyclophilins plays a regulatory role in the heat shock response and the mitochondrial cyclophilin (Cyclophilin D) is an integral part of the mitochondrial permeability transition complex, which is regarded as having a crucial role in mechanisms of cell death. In support of a role in the stress response, the expression of certain cyclophilins has recently been shown to be up‐regulated under various stressful conditions. Current evidence of functional involvement of cyclophilins in various intracellular pathways is reviewed along with the indications that cyclophilin D (Cyp D) represents a crucial part of the mitochondrial permeability transition pore, which is detrimental in apoptotic and necrotic cell death. This review does not attempt to cover all the existing information related to cyclophilin family of proteins, but focus on the existing evidence of the involvement of these proteins in the intracellular stress response.
Age‐related changes in susceptibility of rat brain slice cultures including hippocampus to encephalomyocarditis virus
Authors: Su Weiping , Ueno‐Yamanouchi Aito , Uetsuka Koji , Nakayama Hiroyuki , Doi Kunio
Published: March 2003

Replication of the D variant of encephalomyocarditis virus (EMC‐D) and its cytopathic effects were studied in the brain slice cultures including hippocampus (hippocampal slice) obtained from postnatal 1‐, 4‐, 7‐, 14‐, 28‐and 56‐day‐old Fischer 344 rats. At 0, 12, 24, 36 and 48 h after infection, virus titres of the slices and culture media were assayed. Viral replication was observed in cultures from 1‐to 28‐day‐old rats, and the highest titre was recorded in the slice and culture medium from the youngest rat. The peak of virus titre decreased with age and no distinct viral replication was observed in the cultures from 56‐day‐old rats. Light microscopy revealed that degenerative and necrotic changes appeared in the infected hippocampal slices from 1‐ to 28‐day‐old rats, and the changes became less prominent with age. In situ hybridization and indirect immunofluorescence staining showed that positive signals of viral RNA and antigen were prominent in younger rats and decreased with age. These results suggest that an age‐related decrease in the susceptibility of rat brain to EMC‐D is less related to the maturation of the immune system but possibly to that of the neurone.
Hepatocyte growth factor/scatter factor‐induced intracellular signalling
Authors: Stuart Katherine A. , Riordan Stephen M. , Lidder Sukhwinderjit , Crostella Luca , Williams Roger , Skouteris George G.
Published: March 2003

Hepatocyte growth factor (HGF) identical to scatter factor (SF) is a glycoprotein involved in the development of a number of cellular phenotypes, including proliferation, mitogenesis, formation of branching tubules and, in the case of tumour cells, invasion and metastasis. This fascinating cytokine transduces its activities via its receptor encoded by the c‐met oncogene, coupled to a number of transducers integrating the HGF/SF signal to the cytosol and the nucleus. The downstream transducers coupled to HGF/MET, most of which participate in overlapping pathways, determine the development of the cell's phenotype, which in most cell types is dual.
Tissue granuloma structure‐function in experimental visceral leishmaniasis
Authors: Murray Henry W.
Published: March 2003

In experimental visceral leishmaniasis in normal mice (BALB/c, C57BL/6) acquired resistance to Leishmania donovani, a protozoan which targets tissue macrophages, depends upon T cells, Th1 cell‐type cytokine generation and activated mononuclear phagocytes. In the intact host, initial control and eventual resolution of L. donovani hepatic infection in normal mice is expressed by and accomplished within well‐formed, mature tissue granulomas. In the liver, these immunologically active, inflammatory structures are assembled around a core of fused, parasitized resident macrophages (Kupffer cells) which come to be encircled by both cytokine‐secreting T cells and influxing leishmanicidal blood monocytes. This pro‐host defense granuloma structure‐function relationship, in which histologically mature granulomas provide the microenvironment for intracellular L. donovani killing, however, is only one of seven which have been identified through experimental modifications in this model. This report reviews these structure‐function relationships and illustrates the broad spectrum of additional possible responses. These responses range from structurally intact granulomas which provide no antileishmanial function (the ‘ineffective’ granuloma), to enlarged granulomas which show enhanced parasite killing (the ‘hypertrophied’ granuloma), to effective antileishmanial activity in the absence of any tissue reaction (the ‘invisible’ granuloma).
Alteration in plasma glucose levels in Japanese encephalitis patients
Authors: TANDON APURVA , SINGH ADITI , ATRISHI EKTA , SAXENA S. K. , MATHUR ASHA
Published: March 2003

Summary  A unique factor, human T cell hypoglycaemic factor (hTCHF), has been shown to produce hypoglycaemia during the convalescent stage in the plasma of patients with Japanese encephalitis virus (JEV) infection. The present study was undertaken to investigate the ability of T cells from fresh peripheral blood mononuclear cells (PBMC) of such patients to produce hTCHF. The PBMC, as well as the individual subpopulations, were cultured for 24 h and the culture supernatants (CS) were assayed for hypoglycaemic activity. The activity was observed in the CD8+ T cells. The hypoglycaemia in JE‐confirmed patients coincided with the gradual rise in circulating glucagon level, with no significant alterations in insulin, growth hormone and cortisol levels. The hTCHF was purified by ion exchange chromatography and the purified protein was observed as a ∼25 kDa band on SDS‐PAGE. Secretory hTCHF in the sera of patients and T cell CS was present in 88% of convalescent serum samples. We conclude that during the convalescent stage of JEV infection, a unique factor, hTCHF, is secreted by activated CD8+ T cells from patients and that this is responsible for the development of hypoglycaemia.
Mechanisms of 5‐azacytidine (5AzC)‐induced toxicity in the rat foetal brain
Authors: Ueno Masaki , Katayama Kei‐ichi , Nakayama Hiroyuki , Doi Kunio
Published: March 2003

Mechanisms of 5‐azacytidine (5AzC)‐induced toxicity in the rat foetal brain were investigated. 5AzC (10 mg/kg) was injected into pregnant rats on day 13 of gestation and the protein and mRNA expressions of p53 and its transcriptional target genes, p21, bax, cyclin G1, fas, and gadd45, were examined in the foetal brain. The number of p53‐positive cells peaked at 9 h after treatment (HAT) and those of apoptotic cells and p21‐positive cells peaked at 12 HAT. The expressions of p21, bax, cyclin G1, and fas mRNAs were significantly elevated from 9 to 12 HAT. From the experiments using 5‐bromo‐2′‐deoxyuridine (BrdU), as compared with controls, the migration of neuroepithelial cells significantly delayed and BrdU‐positive signals were observed in many apoptotic cells from 9 to 24 HAT in the 5AzC‐group. In addition, the number of S phase cells significantly decreased at 12 HAT. The present results indicate that 5AzC induced apoptosis and cell cycle arrest probably at G1 phase in the rat foetal brain and they might be mediated by p53 in response to DNA damage.
Fluoride pit and fissure sealants: a review
Authors: Morphis Tonia L. , Toumba Jack K. , Lygidakis Nick A.
Published: March 2003

There are two methods of fluoride incorporation into fissure sealants. In the first method, fluoride is added to the unpolymerized resin in the form of a soluble fluoride salt that releases fluoride ions by dissolution, following sealant application. In the second method, an organic fluoride compound is chemically bound to the resin and the fluoride is released by exchange with other ions (anion exchange system). This report reviews the literature on the effectiveness of all the fluoride‐releasing sealants –commercial and experimental – that have been prepared using either the former or the latter method of fluoride incorporation. There is evidence for equal retention rates to conventional sealants and for ex vivo fluoride release and reduced enamel demineralization. However, further research is necessary to ensure the clinical longevity of fluoride sealant retention and to establish the objective of greater caries inhibition through the fluoride released in saliva and enamel.
Oral health in children undergoing liver transplantation
Authors: Sheehy Evelyn C. , Roberts Graham J. , Beighton David , O’Brien Graham
Published: March 2003

Aims. To determine the prevalence of dental caries in children undergoing liver transplantation and to compare the plaque, gingivitis, and gingival overgrowth indices and oral mucosal lesions in children before and after liver transplantation.

Patients and methods. Patients undergoing liver transplantation at King’s College Hospital were examined before transplantation, at 3 and 106 days post‐transplantation. Healthy children from the Greenwich Health District, south‐east London, were matched to the liver transplant patients by age, gender, socio‐economic factors and the presence or absence of active dental caries.

Results. Twenty‐seven liver transplant patients (mean age: 7 years and 10 months; SD 3 years and 5 months) and 27 controls (mean age: 8 years and 6 months; SD 3 years and 7 months) were examined. The mean dmft and DMFT scores were 2·3 (SD 4·1) and 0·8 (SD 1·4), respectively, in the liver transplant patients, and 1·2 (SD 2·2) and 0·9 (SD 1·5), respectively, in the controls. There were no significant differences between either the mean plaque or gingivitis indices for the primary and permanent teeth in patients before and after transplantation. There were no significant differences between the liver transplant patients and the controls for either mean plaque or gingivitis indices at each examination time. Gingival overgrowth was present in 41% of liver recipients receiving cyclosporin with or without nifedipine, but not in the majority receiving tacrolimus at the final examination. Oral mucosal lesions were absent in both the patients and controls at each examination time.

Conclusions. The oral health of the children undergoing liver transplantation was inadequate. Funding and implementation of an oral health care programme must become a priority for all children before and after liver transplantation.
A three‐dimensional method of visualizing the temporomandibular joint based on magnetic resonance imaging in a case of juvenile chronic arthritis
Authors: Kitai N. , Kreiborg S. , Murakami S. , Bakke M. , Møller E. , Darvann T. A. , Takada K.
Published: March 2003

Summary. Aim. The purpose of this study was to present a new visualizing method for temporomandibular joint (TMJ) pathology in 3D at several condylar positions and to apply the method to a case of juvenile chronic arthritis (JCA) with previously diagnosed bilateral TMJ disorder and to a subject without signs and symptoms from the TMJ.

Subject. The 20‐year‐old female patient had suffered from polyarticular JCA from the age of 6 years 8 months. The present study is based on a follow‐up examination after the completion of orthodontic treatment with the Herbst appliance. Both TMJs were examined using magnetic resonance imaging (MRI) at closed and open mouth positions. The mandibular condyle, the glenoid fossa and the articular disc were segmented and 3D reconstruction of these structures was carried out.

Results. The condyle was characterized by an increased sagittal diameter with osteophyte‐like formation at the anterior aspect of the condylar head and flattening of the superior surface. The depth of the glenoid fossa was reduced and the articular eminence was remarkably flat. The articular disc was markedly diminished and posteriorly placed on the condylar head. The distance of the condylar path during mouth opening was reduced and the curvature of the condylar path was quite flat. The disc moved slightly posteriorly on the condylar head and remained in the glenoid fossa during mouth opening.

Conclusion. The imaging modality described improves visualization of TMJ morphology and gains insight into the TMJ pathology of the JCA patient, adding to understanding of the clinical problems.