Bilateral patellar tendon rupture in a weightlifter during an acute high-loading resistance exercise bout: A case study.

Bilateral patellar tendon ruptures are exceedingly uncommon, especially when they occur in individuals without predisposing risk factors or systemic disease. Due to its rarity, many cases are missed on initial presentation resulting in poor patient outcomes. Identifying associated risk factors aids in diagnosis and mitigates this oversight. We report a case of a healthy, recreational weightlifter who sustained bilateral patellar tendon ruptures during an acute high-loading resistance exercise bout. We discuss how a spike in acute workload may have predisposed our patient to this injury. Research into training load and athlete injury risk is currently in vogue, however, no studies have analysed whether poor load management increases the risk of tendon ruptures. This case prompts awareness for clinicians who diagnose and manage this injury and helps to stimulate the formation of educational initiatives for athletes and coaches, aimed at injury prevention.

Presenting features of female collegiate sports-related concussion in South Africa: a descriptive analysis.

Background: Sports-related concussion (SRC) is an injury with important implications, especially in collision and contact sports, and has a high symptom burden. Student athletes face particular psychosocial challenges, especially female students with pre-existing anxiety/depression are at increased risk for SRC, and have a higher symptom burden before and after injury. Objectives: Describing female SRC presenting features at a collegiate campus-based sports medicine service; examining the association of prior concussion history (PCONC) and pre-existing anxiety/depression (PMHDx) with SRC. Methods: A retrospective cohort and statistical analysis (including corrected effect sizes) of Sport Concussion Assessment Tool (versions 3/5) data (Step 1: PCONC and PMHDx history; Step 2: symptom evaluation) of collegiate female athletes with SRC between 2012 and 2018. Results: Forty females with SRC were identified (age 23 ± 3). The five most frequent symptoms were headache (n = 34; 85%), feeling slowed down (n = 33; 83%), pressure in head (n = 33; 83%), don't feel right (n = 32; 80%) and fatigue/low-energy (n = 32; 80%). These five symptoms also had the highest self-rated severity (median (IQR): headache (3 (2-4)), feeling slowed down (3 (1-4)), fatigue/low-energy (3 (1-5)), don't feel right (3 (1-4)) and pressure in head (3 (2-4)). PMHDx (n = 8; 62.9 vs 38.6; p = 0.0192; Hedges' gs = 0.95; large ES), and not PCONC (n = 13; 51.0 vs 39.8; p = 0.2183; Hedges' gs = 0.48; small ES) was associated with increased mean total symptom severity. Conclusion: Headache, feeling slowed down, pressure in head, don't feel right and fatigue/low-energy had the highest symptom burden. Total symptom severity was no different in those with and without PCONC, but significantly higher in those with PMHDx.

Skeletal muscle telomere length in healthy, experienced, endurance runners.

Measuring the DNA telomere length of skeletal muscle in experienced endurance runners may contribute to our understanding of the effects of chronic exposure to endurance exercise on skeletal muscle. This study compared the minimum terminal restriction fragment (TRF) length in the vastus lateralis muscle of 18 experienced endurance runners (mean age: 42 +/- 7 years) to those of 19 sedentary individuals (mean age: 39 +/- 10 years). The runners had covered almost 50,000 km in training and racing over 15 years. Minimum TRF lengths measured in the muscle of both groups were similar (P = 0.805) and within the normal range. Minimum TRF length in the runners, however, was inversely related to their years spent running (r = -0.63, P = 0.007) and hours spent training (r = -0.52, P = 0.035). Therefore, since exposure to endurance running may influence minimum TRF length, and by implication, the proliferative potential of the satellite cells, chronic endurance running may be seen as a stressor to skeletal muscle.

5,6-Dichloro-1-Beta-D-ribofuranosylbenzimidazole inhibits initiation of nuclear heterogeneous RNA chains in HeLa cells.

The nucleoside analog 5, 6-dichloro-1-beta-d-ribofuranosylbenzimidazole (DRB) at 75 to 150 micromolar concentrations inhibits the synthesis of nuclear heterogeneous RNA (hnRNA) in HeLa cells by 60 to 70 percent. The sedimentation profile of hnRNA labeled with (3H)uridine for 45 seconds after brief treatment (45, 90, or 180 seconds) with DRB showed a progressive decrease in the labeling of shorter hnRNA molecules relative to longer molecules. Prior exposure of the cells to actinomycin D, an inhibitor of RNA chain elongation, did not alter the sedimentation profile of hnRNA. These results suggest that DRB preferentially inhibits the initiation of hnRNA chains so that after exposure to DRB for a brief period the longer nascent chains still remain to be finished and thus incorporate a greater share of the pulse label. By progressively increasing the time of exposure to DRB, and measuring the rate of increase in the average size of the labeled, nascent RNA, it was estimated that the chains were growing at rates between 50 and 100 nucleotides per second.

Silent and expressed sister Mup genes are located within distinct chromatin domains: analysis by pulsed-field gel electrophoresis and polymerase chain reaction-supplemented DNase I digestion.

We have recently described a subfamily of two genes, Mup-1.5a and Mup-1.5b, which exist as a nonallelic pair in most inbred strains of mice. The Mup-1.5a and Mup-1.5b genes are more than 99.9% homologous, yet they are differentially expressed. While the Mup-1.5a gene is expressed at a high level in the submaxillary gland, the Mup-1.5b gene does not appear to be expressed either in this or in any other tissue. The Mup-1.5b gene can, however, be expressed as a transgene with the tissue specificity of its sister gene, Mup-1.5a. We have shown before that both the Mup-1.5a and Mup-1.5b genes are located on chromosome 4, closely linked to the Mup-1 locus. In this report, we demonstrate the two genes are located within distinct chromosomal domains, separated by at least 150 to 200 kb of DNA. Using a novel method, detailed in this report, we show that in the submaxillary gland, the Mup-1.5a gene is five- to sixfold more susceptible to DNase I digestion than is the Mup-1.5b gene. This finding suggests that the inactivity of the Mup-1.5b gene is brought about by long range-acting mechanisms that establish a chromatin structure in the vicinity of this gene incompatible with transcription.

Tissue-specific expression of major urinary protein (MUP) genes in mice: characterization of MUP mRNAs by restriction mapping of cDNA and by in vitro translation.

The major urinary proteins (MUPs) in mice are coded for by a gene family which consists of ca. 30 members. The number of MUP genes that are expressed is not known. Previous studies have shown that MUP mRNAs are present in several secretory tissues in addition to the liver, in which they were originally identified. In this paper we show, through restriction analysis of MUP cDNAs, that distinct sets of MUP mRNAs are synthesized in each of the tissues studied and that these mRNAs are most likely coded for by different genes. As is shown, MUP mRNAs of different tissues are related to an extent that precludes the use of gene-specific probes in differentiating among them. The regions of homology also include the 3' untranslated regions of MUP mRNAs. The question of differential expression was thus investigated by searching for restriction polymorphisms in MUP mRNAs. We demonstrate that subtle differences in the sequences of even scarce mRNAs can be recognized by this particular approach. In addition, it is shown that MUP mRNAs of different tissues code for different, nonoverlapping sets of polypeptides, as determined by gel electrophoresis of in vitro-translated precursors to MUPs. The relevance of these results to models of evolution of tissue-specific regulation in a multigene family is discussed.

Nucleotide sequences of liver, lachrymal, and submaxillary gland mouse major urinary protein mRNAs: mosaic structure and construction of panels of gene-specific synthetic oligonucleotide probes.

The mouse major urinary proteins (MUPs) are encoded by a gene family of about 35 to 40 members. MUPs are synthesized in at least six secretory tissues under a variety of developmental and endocrine controls, but the identities of the individual genes expressed in each tissue have not previously been established. In this article, we present the nucleotide sequences of five MUP mRNAs which we designate MUP I through V. MUPs I, II, and III are the most abundant MUP mRNA species in the liver, and MUPs IV and V are the most abundant MUP mRNA species in the lachrymal gland and the submaxillary gland, respectively. The sequence data show that each of the five mRNAs is encoded by a distinct member of the gene family. The structures of the MUP mRNA consist of interspersed segments of variable and conserved sequences. On the basis of the sequences of the variable segments, gene-specific panels of synthetic oligonucleotide probes were prepared. The gene-specific panels were used to identify cloned genes and, as described in the accompanying paper (K. Shahan, M. Denaro, M. Gilmartin, Y. Shi, and E. Derman, Mol. Cell. Biol. 7:1947-1954, 1987), to characterize the expression of MUP genes I through V.

Identification of an enhancer required for the expression of a mouse major urinary protein gene in the submaxillary gland.

The MUP1.5b gene was previously found to be expressed specifically in the submaxillary gland and at high levels when introduced into mice as a transgene including 4.7 kb of 5'-flanking DNA and 0.3 kb of 3'-flanking DNA. To localize regulatory elements responsible for this tissue-specific pattern of expression, we tested the expression of three additional MUP1.5b transgenes including various amounts of 5'-flanking DNA. These experiments indicated that sequences between -1.85 and -3.46 kb from the transcription initiation site were required for high-level expression in the submaxillary gland. The presence of regulatory elements in this region was also suggested by the detection of a DNase I-hypersensitive site, seen only in submaxillary gland nuclei, at position -2.5 kb upstream from the MUP1.5a gene, a member of the same MUP gene subfamily and virtually identical to the MUP1.5b gene. Further evidence for enhancer activity was provided by the ability of the 1.6-kb DNA fragment including sequences between -1.85 and -3.46 kb to stimulate the expression of an otherwise inactive MUP1.5b-chloramphenicol acetyltransferase fusion gene specifically in the submaxillary gland. The nucleotide sequence of this 1.6-kb DNA fragment was found to be identical for the MUP1.5a and MUP1.5b genes. Together, these results provide the first localization of a cis-acting regulatory sequence involved in the differential tissue-specific expression of the MUP gene family.

Expression of six mouse major urinary protein genes in the mammary, parotid, sublingual, submaxillary, and lachrymal glands and in the liver.

Mouse major urinary proteins (MUPs) are encoded by a family of about 35 to 40 highly conserved genes. In the preceding paper (K. Shahan, M. Gilmartin, and E. Derman, Mol. Cell. Biol. 7:1938-1946, 1987), we presented the sequences of the most abundant MUP mRNAs in the liver (MUP I, II, and III) and in the lachrymal (MUP IV) and submaxillary (MUP V) glands. We have shown that these five mRNAs are coded by five distinct genes, MUP I through V. In the present communication, we examine the expression of MUP genes in all of the six tissues in which MUP mRNAs are synthesized, the mammary, parotid, sublingual, lachrymal, and submaxillary glands and the liver. We show that gene MUP II is expressed in the liver and in the mammary gland, that gene MUP IV is expressed in the lachrymal and parotid glands, and that gene MUP V is expressed in the submaxillary, sublingual, and lachrymal and parotid glands, and that gene MUP V is expressed in the submaxillary, sublingual, and lachrymal glands. Furthermore, we present evidence that in addition to genes MUP I through V, another gene, MUP VI, is expressed in BALB/c mice in the parotid gland. The tissue-specific synthesis of MUP mRNAs is thus brought about by two major mechanisms: the expression, in different tissues, of different members of the family and the expression of a single gene at various levels in different tissues. When a particular MUP gene is expressed in several tissues, transcripts of this gene initiate at the same site and are spliced and polyadenylated in the same manner.

Athletes with exercise-associated fatigue have abnormally short muscle DNA telomeres.

INTRODUCTION/PURPOSE: Although the beneficial health effects of regular moderate exercise are well established, there is substantial evidence that the heavy training and racing carried out by endurance athletes can cause skeletal muscle damage. This damage is repaired by satellite cells that can undergo a finite number of cell divisions. In this study, we have compared a marker of skeletal muscle regeneration of athletes with exercise-associated chronic fatigue, a condition labeled the "fatigued athlete myopathic syndrome" (FAMS), with healthy asymptomatic age- and mileage-matched control endurance athletes. METHODS: Muscle biopsies of the vastus lateralis were obtained from 13 patients diagnosed with FAMS and from 13 healthy control subjects. DNA was extracted from the muscle samples and their telomeric restriction fragment (TRF) or telomere lengths were measured by Southern blot analysis. RESULTS: All 13 symptomatic athletes reported a progressive decline in athletic performance, decreased ability to tolerate high mileage training, and excessive muscular fatigue during exercise. The minimum value of TRF lengths (4.0 +/- 1.8 kb) measured on the DNA from vastus lateralis biopsies from these athletes were significantly shorter than those from 13 age- and mileage-matched control athletes (5.4 +/- 0.6 kb, P < 0.05). Three of the FAMS patients had extremely short telomeres (1.0 +/- 0.3 kb). The minimum TRF lengths of the remaining 10 symptomatic athletes (4.9 +/- 0.5 kb, P < 0.05) were also significantly shorter that those of the control athletes. CONCLUSION: These findings suggest that skeletal muscle from symptomatic athletes with FAMS show extensive regeneration which most probably results from more frequent bouts of satellite cell proliferation in response to recurrent training- and racing-induced muscle injury.

[The mathematical modelling of the displacement of the hard palate tissues in conservative uranoplasty]. / Matematicheskoe modelirovanie peremeshcheniia tkanei tverdogo neba pri shchadiashchei uranoplastike.

Presents a mathematical model of sparing uranoplasty techniques developed by the authors; using this model, a physician may assess before surgery the volume of tissues needed to close the palatal defect. Experience gained with 72 surgeries carried out in children with unilateral cleft palate confirms the desirability of preliminary computations. Mathematical models for the most prevalent sparing uranoplasty methods are offered.

Peripheral arterial disease and intermittent claudication: efficacy of short-term upper body strength training, dynamic exercise training, and advice to exercise at home.

OBJECTIVE: To compare the effect of two training programmes and advice to exercise at home on physiological adaptations in patients with peripheral arterial disease (PAD). DESIGN: 30 patients with a typical history of PAD and intermittent claudication were randomised to either an upper body strength training programme (UBST), a dynamic (walking, cycling, circuit) conventional exercise rehabilitation programme (CER), or advice to 'walk as much as possible at home' (CONT). Before and after intervention groups performed a standard graded treadmill exercise test (GTET) and a 6-minute walk test (SMWT) to determine peak physiological parameters and walking distances. Maximal walking distance (MWD), pain-free walking distance (PFWD), peak oxygen uptake (VO2) , heart rate and perceived pain were measured. RESULTS: MWD on the GTET increased significantly in the CER group compared with the CONT and UBST groups (93.9 +/- 79% v. 7.0 +/- 19.8% v. 7.3 +/- 46%; CER v. UBST v. CONT p = 0.003). Similarly, peak VO2 increased with CER compared with the CONT and UBST groups (28.4 +/- 20 v. -6.2 +/- 15 v. -1.0 +/- 21%; CER v. UBST v. CONT p = 0.004). During the SMWT the CER and UBST groups improved in PFWD compared with the CONT group (37 +/- 47% v. 27 +/- 71% v. -30 +/- 29%; CER v. UBST v. CONT p = 0.03), and perceived pain decreased in the CER group compared with the UBST group (-24 +/- 39% v. 27 +/- 48%; CER v. UBST p = 0.01). CONCLUSION: CER improves physiological parameters and walking distances more than UBST does. CER is effective within 6 weeks. Verbal encouragement to exercise is an ineffective form of management.

Factors predicting walking intolerance in patients with peripheral arterial disease and intermittent claudication.

OBJECTIVE: To determine which physiological variables conduce to walking intolerance in patients with peripheral arterial disease (PAD). DESIGN: The physiological response to a graded treadmill exercise test (GTT) in patients with PAD was characterised. SETTING: Patients were recruited from the Department of Vascular Surgery, Groote Schuur Hospital, Cape Town. SUBJECTS: Thirty-one patients diagnosed with PAD were included in the study. OUTCOME MEASURES: During a GTT, peak oxygen consumption (VO(2peak)), peak minute ventilation (VE(peak)), peak heart rate and peak venous lactate concentrations were measured and compared with those from a comparison group. Ankle-brachial index (ABI) was measured at rest and after exercise. During the GTT, maximum walking distance (MWD) and pain-free walking distance (PFWD) were measured to determine walking tolerance. RESULTS: Peak venous lactate concentrations did not correlate significantly with either PFWD (r = -0.08; p = 0.3) or MWD (r = -0.03; p = 0.4). Resting ABI did not correlate with either MWD (r = 0.09; p = 0.64) or PFWD (r = -0.19; p = 0.29). Subjects terminated exercise at significantly (p < 0.05) lower levels of cardiorespiratory effort and venous lactate concentrations than did a sedentary but otherwise healthy comparison group: peak heart rate 156 +/- 11 v. 114 +/- 22 beats per minute (BPM); p = 0.001; and peak venous lactate concentration 9.7 +/- 2.7 mmol/l v. 3.28 +/- 1.39 mmol/1; p = 0.001. CONCLUSION: Perceived discomfort in these patients is not caused by elevated blood lactate concentrations, a low ABI or limiting cardiorespiratory effort but by other factors not measured in this study.

International Olympic Committee consensus statement: molecular basis of connective tissue and muscle injuries in sport.

Tendon and ligament injures cause significant loss of performance in sport and decreased functional capacity in the workplace. Many of these injures remain difficult to treat, and many individuals have long-term pain and discomfort. Animal studies of growth factor and cell-based therapies have shown promising results, but these treatments also can be misused to enhance athletic performance. The International Olympic Committee (IOC) now has high-level scientific advisors who can advise the IOC as to the use and abuse of these technologies.

Evaluation of maximal exercise performance, fatigue, and depression in athletes with acquired chronic training intolerance.

OBJECTIVE: This study compared differences in maximal strength and aerobic capacity and symptoms of fatigue and depression in athletes with acquired training intolerance (ATI) and control athletes (CON) matched for age and current training volume who did not have symptoms of excessive or chronic fatigue associated with their sporting activity. SETTING: University of Cape Town, Sports Science Institute of South Africa. PARTICIPANTS: Twenty ATI and 10 CON athletes participated in the trial. Although the ATI athletes reported symptoms of excessive fatigue during exercise, or symptoms of fatigue that occurred at rest and during activities of daily living, they did not fulfill the criteria for a diagnosis of chronic fatigue syndrome. MAIN OUTCOME MEASURES: A training and comprehensive medical history was recorded from all subjects. The Beck Depression Inventory Short Form (BDI-SF) was used to assess levels of depression in both ATI and control subjects. Maximal force output during a 5-second isometric voluntary knee extensor muscle contraction, and maximal aerobic capacity (VO2max), maximal heart rate (HRmax), and maximal blood lactate concentrations during a treadmill running test were measured in all subjects. RESULTS: There were no differences in maximal isometric force output, peak treadmill running speed, VO2max, HRmax, or blood lactate concentration at rest or after maximal exercise testing between the ATI and CON athletes. However, the BDI-SF scores were higher in the ATI (7.7 +/- 6.6 arbitrary units) than in the CON athletes (1.7 +/- 1.5 arbitrary units; (P = 0.0052). CONCLUSIONS: These findings suggest that the symptoms of excessive fatigue and acquired training intolerance described by these ATI athletes do not affect their maximal isometric and maximal aerobic capacity, and may be associated with psychologic depression in these athletes.

Isolation of a cDNA clone for mouse urinary proteins: age- and sex-related expression of mouse urinary protein genes is transcriptionally controlled.

A recombinant cDNA plasmid derived from mouse urinary protein (MUP) mRNA was isolated and used to determine the level of control of the developmentally regulated and the sex-linked expression of MUP genes by monitoring the transcription of MUP mRNA sequences in isolated liver nuclei. No transcription of MUP genes could be detected in liver nuclei of prepubescent animals whose livers do not contain measurable MUP mRNA. Transcription of MUP genes in the livers of adult male mice was 6-fold higher than in the livers of adult female mice, proportional to the difference in MUP mRNA concentrations. Transcriptional control mechanisms are therefore implicated as responsible for both the developmentally and the sex-linked changes in the expression of MUP genes.

Aetiology of skeletal muscle 'cramps' during exercise: a novel hypothesis.

The aetiology of exercise-associated muscle cramps (EAMC), defined as 'painful, spasmodic, involuntary contractions of skeletal muscle during or immediately after physical exercise', has not been well investigated and is therefore not well understood. This review focuses on the physiological basis for skeletal muscle relaxation, a historical perspective and analysis of the commonly postulated causes of EAMC, and known facts about EAMC from recent clinical studies. Historically, the causes of EAMC have been proposed as (1) inherited abnormalities of substrate metabolism ('metabolic theory') (2) abnormalities of fluid balance ('dehydration theory'), (3) abnormalities of serum electrolyte concentrations ('electrolyte theory') and (4) extreme environmental conditions of heat or cold ('environmental theory'). Detailed analyses of the available scientific literature including data from recent studies do not support these hypothesis for the causes of EAMC. In a recent study, electromyographic (EMG) data obtained from runners during EAMC revealed that baseline activity is increased (between spasms of cramping) and that a reduction in the baseline EMG activity correlates well with clinical recovery. Furthermore, during acute EAMC the EMG activity is high, and passive stretching is effective in reducing EMG activity. This relieves the cramp probably by invoking the inverse stretch reflex. In two animal studies, abnormal reflex activity of the muscle spindle (increased activity) and the Golgi tendon organ (decreased activity) has been observed in fatigued muscle. We hypothesize that EAMC is caused by sustained abnormal spinal reflex activity which appears to be secondary to muscle fatigue. Local muscle fatigue is therefore responsible for increased muscle spindle afferent and decreased Golgi tendon organ afferent activity. Muscles which cross two joints can more easily be placed in shortened positions during exercise and would therefore decrease the Golgi tendon organ afferent activity. In addition, sustained abnormal reflex activity would explain increased baseline EMG activity between acute bouts of cramping. Finally, passive stretching invokes afferent activity from the Golgi tendon organ, thereby relieving the cramp and decreasing EMG activity.

A lumbar body support (KBS 2000) alters lumbar muscle recruitment patterns in patients with acute-upon-chronic lower back pain.

OBJECTIVE: To determine the effects of a locally designed lumbar body support (LBS) on integrated electromyographic (IEMG) activity of the lumbar erector spinae muscles, on heart rate and on ratings of discomfort in patients with low back pain. DESIGN: Non-randomised controlled trial. SETTING: Patients referred from general practitioners and back pain rehabilitation programmes in Cape Town. PATIENTS AND OTHER PARTICIPANTS: Ten patients with low back pain of diverse causes. Values were compared with those in 10 control subjects without low back pain. INTERVENTION: Patients and controls lay supine on (in random order) either a flat conventional mattress or a LBS placed on top of the flat mattress, for a 30-minute period (acute exposure), and every night for 2 weeks (chronic exposure). MAIN OUTCOME MEASURES: IEMG activity of the lumbar erector spinae muscles, heart rate, and perception of comfort. RESULTS: IEMG activity of the lumbar erector spinae muscles did not differ between controls and patients when lying on the LBS on top of the CM after either acute or chronic exposure. However, it was significantly greater (P < 0.05) in patients than in controls when lying on the flat mattress. Subjective ratings of discomfort and heart rates mirrored these changes and were higher in patients only when lying on the flat mattress (P < 0.05). Patients with low back pain also reported that sleeping overnight on the LBS on top of their own mattress significantly reduced discomfort ratings. CONCLUSIONS: When lying on a mattress with a flat surface, patients with chronic low back pain have higher IEMG activity of the erector spinae muscles, higher heart rates and higher subjective ratings of discomfort than do control subjects. These differences disappear when both groups use a specially designed lumbar body support placed on top of the flat surface. These preliminary studies suggest that a lumbar body support should be evaluated in the chronic management of low back pain.

hnRNA in HeLa cells: distribution of transcript sizes estimated from nascent molecule profile.

In HeLa cells, about half the hnRNA is synthesized from transcription units which are less than 5000 bases in length, although over 80% is larger than the average mRNA. This conclusion was derived from a quantitative analysis of the nascent chain sedimentation profile.

Subfamily of submaxillary gland-specific Mup genes: chromosomal linkage and sequence comparison with liver-specific Mup genes.

Mouse major urinary proteins (MUPs) are encoded by a family of ca. 35 genes that are expressed in a tissue-specific manner in several secretory organs; in the liver, in the submaxillary, sublingual, parotid and lachrymal glands, and in the skin sebaceous glands. In this paper we describe the isolation of a Mup gene, Mup-1.5a, which is expressed predominantly in the submaxillary gland of BALB/c mice. We show that Mup-1.5a is a member of a subfamily consisting of two closely related genes, both of which are closely linked to the Mup-1 locus on mouse chromosome 4. Mup-1 is the locus of a class of Mup genes (Group 1) expressed in the liver. The complete nucleotide sequence of Mup-1.5a has been determined, and was compared to a previously sequenced Group 1 Mup gene. The comparison shows that the differentially expressed Mup genes are uniformly divergent in exons, introns and in their flanking sequences. The regions of homology extend at least 5 kb into the 5' flanking region of Mup genes.