Inducement of tissue regeneration of harvested hamstring tendons in a rabbit model.

OBJECTIVES: The objective of this study was to determine if the use of fascia lata as a tendon regeneration guide (placed into the tendon canal following harvesting the semitendinosus tendon) would improve the incidence of tissue regeneration and prevent fatty degeneration of the semitendinosus muscle. MATERIALS AND METHODS: Bilateral semitendinosus tendons were harvested from rabbits using a tendon stripper. On the inducing graft (IG) side, the tendon canal and semitendinosus tibial attachment site were connected by the fascia lata, which was harvested at the same width as the semitendinosus tendon. On the control side, no special procedures were performed. Two groups of six rabbits were killed at post-operative weeks 4 and 8, respectively. In addition, three healthy rabbits were killed to obtain normal tissue. We evaluated the incidence of tendon tissue regeneration, cross-sectional area of the regenerated tendon tissue and proportion of fatty tissue in the semitendinosus muscle. RESULTS: At post-operative week 8, the distal end of the regenerated tissue reached the vicinity of the tibial insertion on the control side in two of six specimens. On the IG side, the regenerated tissue maintained continuity with the tibial insertion in all specimens. The cross-sectional area of the IG side was significantly greater than that of the control side. The proportion of fatty tissue in the semitendinosus muscle on the IG side was comparable with that of the control side, but was significantly greater than that of the normal muscle. CONCLUSIONS: Tendon tissue regenerated with the fascia lata graft was thicker than naturally occurring regenerated tissue. However, the proportion of fatty tissue in the semitendinosus muscle was greater than that of normal muscle.Cite this article: K. Tabuchi, T. Soejima, H. Murakami, K. Noguchi, N. Shiba, K. Nagata. Inducement of tissue regeneration of harvested hamstring tendons in a rabbit model. Bone Joint Res 2016;5:247-252. DOI: 10.1302/2046-3758.56.2000585.

Effect of aging on volatile compounds in cooked beef.

Volatiles in the headspace of beef cooked at 180 °C were analyzed using solid-phase microextraction (SPME) and gas chromatography-mass spectrometry (GC-MS), and the effects of aging were evaluated. Seventy volatile substances including non-aromatic, homocyclic, and heterocyclic compounds were identified. A significant positive regression model for storage could be adopted for toluene, benzeneacetaldehyde, 2-formylfuran, pyrazine, 2,6-dimethylpyrazine, 2,3-dimethylpyrazine, 2-acetylthiazole, and 2-formyl-3-methylthiophene. Increases in the quantity of these compounds, with the exception of toluene, suggest the importance of the Strecker and Maillard reactions in cooked meat previously aged under vacuum conditions. As such, the aging process may lead to an increase not only in the amount of compounds related to the taste of meat, but also in the quantity of odor-active compounds. The increased quantity of toluene during storage seemed to be influenced by lipid oxidation.

High expression of EVI1 and MEL1 is a compelling poor prognostic marker of pediatric AML.

EVI1 and MEL1 are homolog genes whose transcriptional activations by chromosomal translocations are known in small subsets of leukemia. From gene expression profiling data of 130 Japanese pediatric acute myeloid leukemia (AML) patients, we found that EVI1 and MEL1 were overexpressed in ~30% of patients without obvious translocations of these gene loci, and that their high expression was significantly associated with inferior survival. High EVI1 expression was detected mainly in myelomonocytic-lineage (designated as e-M4/M5 subtype) leukemia with MLL rearrangements and in megakaryocytic-lineage (designated as e-M7 subtype) leukemia, and its prognostic association was observed in the e-M4/M5 subtype but not in the e-M7 subtype. On the other hand, high MEL1 expression was detected in myelocytic-lineage (designated as e-M0/M1/M2 subtype) and e-M4/M5 subtype leukemia without MLL rearrangements, and its prognostic association was independent from the subtypes. Because of their subtype-dependent and mutually exclusive expression, a combined evaluation of their high expression enabled a clear distinction of patients with inferior survival (P<0.00001 in event-free survival (EFS) and overall survival (OS)). This association was confirmed by quantitative reverse transcription PCR analysis of an independent cohort of 81 patients (P=0.00017 in EFS, P=0.00028 in OS). We propose that the combined estimation of EVI1 and MEL1 expression will be an effective method to predict the prognosis of pediatric AML.

A unique case of segmental vasal atresia.

We report a case of a 2-year-old boy who presented with an empty left scrotum. Clinical examination revealed a left palpable undescended testis. During orchidopexy, segmental atresia of the vas deferens was found, and microsurgical repair was carried out. Segmental vasal atresia is an extremely rare condition and is infrequently diagnosed, especially in the pediatric age group. This is the first reported case of segmental vasal atresia without an association with cystic fibrosis.

Discrete element analysis in musculoskeletal biomechanics.

This paper is written to honor Professor Y. C. Fung, the applied mechanician who has made seminal contributions in biomechanics. His work has generated great spin-off utility in the field of musculoskeletal biomechanics. Following the concept of the Rigid Body-Spring Model theory by T. Kawai (1978) for non-linear analysis of beam, plate, and shell structures and the soil-gravel mixture foundation, we have derived a generalized Discrete Element Analysis (DEA) method to determine human articular joint contact pressure, constraining ligament tension and bone-implant interface stresses. The basic formulation of DEA to solve linear problems is reviewed. The derivation of non-linear springs for the cartilage in normal diarthrodial joint contact problem was briefly summarized. Numerical implementation of the DEA method for both linear and non-linear springs is presented. This method was able to generate comparable results to the classic contact stress problem (the Hertzian solution) and the use of Finite Element Modeling (FEM) technique on selected models. Selected applications in human knee and hip joints are demonstrated. In addition, the femoral joint prosthesis stem/bone interface stresses in a non-cemented fixation were analyzed using a 2D plane-strain approach. The DEA method has the advantages of ease in creating the model and reducing computational time for joints of irregular geometry. However, for the analysis of joint tissue stresses, the FEA technique remains the method of choice.

Effects of alpha-tocopherol on lactones in beef headspace during storage.

We studied the effect of alpha-tocopherol (alpha-Toc) on the production of lactones, a desirable flavor in Wagyu beef (Japanese Black Cattle), in meat headspace. Fresh beef was minced and divided into 4 groups containing different alpha-Toc levels (1.9, 2.7, 6.1, and 10.7 mg/kg) or packed with oxygen absorber. After 10 d of storage at 2 °C, these samples were cooked in distilled water and rendered fat was collected. The headspace of the fat was analyzed with solid-phase microextraction combined with gas chromatography-mass spectrometry. The DB-17MS capillary column led to good separation of aliphatic lactones. Oxygen was important for the development of all lactones during storage. The amounts of gamma-octalactone and gamma-nonalactone decreased linearly with increasing alpha-Toc levels. The effect of alpha-Toc on delta-decalactone, delta-undecalactone, and delta-tetoradecalactone was weaker, and that on delta-hexadecalactone was ambiguous. These observations suggest that oxidation was necessary to develop the flavor of Wagyu beef, and warn that an excessive increase of alpha-Toc could reduce levels of positive flavor components, gamma-octalactone and gamma-nonalactone, in the headspace of Wagyu beef.

Splenogonadal fusion.

We report a case of an 11-year-old boy who presented with a painless left scrotal mass of one year's duration. Clinical examination and diagnostic modalities indicated a solid left testicular mass. Exploration was done and a complete splenogonadal fusion was found. The ectopic splenic tissue was completely removed with preservation of the testis. Splenogonadal fusion is a rare and infrequently reported entity in the paediatric surgical literature. This was our first encounter with this condition. A review of the literature is briefly presented here.

Analysis of volatile compounds in beef fat by dynamic-headspace solid-phase microextraction combined with gas chromatography-mass spectrometry.

A solid-phase microextraction (SPME) technique has been applied to the determination of the volatile compounds, including diterpenoids and lactones, in cooked beef fat. The ability of static-headspace SPME to extract lactones was disappointing, regardless of the type of SPME fiber or the temperature used. Dynamic-headspace SPME extraction with 50-/30-microm divinylbenzene-Carboxen on a polydimethylsiloxane fiber at 100 degrees C, by contrast, enabled the analysis of volatiles, including delta-lactones, gamma-lactones, and diterpenoids, with 50-/30-microm divinylbenzene-Carboxen on a polydimethylsiloxane fiber at 100 degrees C. Fifty-three compounds were identified from only 0.20 g of rendered beef fat, and 76% of these showed reliable peak size repeatability: the coefficient of variation was less than 10% on the total ion chromatograms obtained from gas chromatography-mass spectrometry (GC-MS) analysis. Some lactones showed higher CV values (>10%), but single-ion mode GC-MS analysis reduced them to 10% or less. In a study of beef samples available to the Japanese market, our analytical procedure revealed significantly higher levels of 1-hexanol, octadecane, ethyl tetradecanoate, gamma-nonalactone, but lower levels of delta-decalactone, delta-dodecalactone, and neophytadiene, in Japanese Black cattle than in beef imported from Australia.

Histochemical properties and collagen architecture of M. iliotibialis lateralis and M. puboischiofemoralis in male broilers with different growth rates induced by feeding at different planes of nutrition.

1. The histochemical properties and the collagen content and architecture of the iliotibialis lateralis (ITL) and puboischiofemoralis (PIF) muscles were assessed in Red Cornish x New Hampshire cockerels reared on a high nutrient plane for 80 d (H80d), or a low nutrient plane for 80 d (L80d) or 95 d (L95d). 2. Final live weights were 3410 g in H80d, 2810 g in L80d and 3467 g in L95d. Both ITL and PIF muscle weights were lowest in L80d and did not differ between H80d and L95d. 3. ITL muscle was composed of fast-twitch myofibres such as IIA (high reduced nicotinamide adenine dinucleotide dehydrogenase, NADH-DH activity), IIB (low NADH-DH activity) and IIC (intermediate NADH-DH activity). The high percentage of type IIB myofibres in H80d (76.6%) and L95d (76.2%) birds were reflected in low percentages of type IIC myofibres (12.2%) in H80d birds and type IIA myofibres (8.2%) in L95d birds. Percentages of IIA, IIB and IIC myofibres in L80d cockerels were 12.4, 69.8 and 17.6%, respectively. 4. The myofibres in PIF muscle were divided into two basic types, I and IIA, and a transitional form (I-tr) from IIA to I. In the caudal region, all myofibres in H80d and L95d cockerels were type I but in L80d cockerels 15% of myofibres were categorised as type I-tr. In the cranial region, the great majority (52 to 63%) of myofibres were type IIA. Type I myofibres occurred at a higher percentage in H80d (30.5%) than L95d (21.8%) and type I-tr in L95d (15.7%) than H80d (7.3%) and L80d (11.5%). 5. The total amount of collagen was higher in ITL than PIF muscle in every bird group. In both muscles the highest collagen content was in L95d cockerels but the content did not differ between H80d and L80d birds. The thickness of thick and thin perimysia increased with muscle size. The circular collagen fibre in the thick perimysium was larger in ITL (6.1 to 7.0 microm) than PIF (3.7 to 3.8 microm) muscle but did not differ among the bird groups. 6. From these results, it was concluded that feeding on a high nutritional plane promotes growth of the thigh muscles, with accompanying enlargement of the perimysial thickness, no increase in collagen content and various changes of histochemical properties.

Effects of nutritional level on muscle development, histochemical properties of myofibre and collagen architecture in the pectoralis muscle of male broilers.

1. The effects of nutritional level on muscle development, histochemical properties of myofibre and collagen architecture in the pectoralis muscle were evaluated using male broilers of Red Cornish x New Hampshire stock, reared on diets of high nutritional value for up to 80 d (H80d) and low nutritional value for up to 80 d (L80d, same age as H80d) or 95 d (L95d, same body weight as H80d). 2. The total live weight and the weight of pectoralis muscle were lower in L80d than in both H80d and L95d. The muscle weight as a percentage of live weight was 8.7% in L80d, 10.7% in H80d and 11.5% in L95d. 3. Pectoralis muscle was composed only of type IIB myofibres and showed no differences in myofibre type composition among the chicken groups. The largest diameter of type IIB myofibres was observed in L95d, followed by H80d and the smallest in L80d. 4. The total amount of intramuscular collagen did not differ among the chicken groups (1.92 to 1.99 mg/g). Types I and III collagens were immunohistochemically detected in both the perimysia and endomysia. The thin perimysia around the primary myofibre fascicles showed larger width in H80d than L80d and L95d, and also the thick perimysia around the secondary fascicles in H80d than L80d. 5. The collagen structure of the perimysium was most developed in H80d, followed by L95d and on the least in L80d. The development of perimysial collagen fibres could be enhanced by a rapid growth rate of the muscle induced by high nutritional level and depressed by a slow growth rate with low nutritional foods. 6. The endomysial collagen architecture was observed as a felt-like tissue of the fibril bundles with many slits. The thinnest endomysial wall was observed in L80d, followed by H80d and the thickest in L95d. 7. From these results, it was indicated that foods of high nutritional value could enhance growth of the pectoralis muscle of broilers, and this is accompanied by hypertrophy of the type IIB myofibres and development of the perimysial collagen architecture.

Developmental states of the collagen content, distribution and architecture in the pectoralis, iliotibialis lateralis and puboischiofemoralis muscles of male Red Cornish x New Hampshire and normal broilers.

1. Developmental states of the collagen content, distribution and architecture in the pectoralis (PT), iliotibialis lateralis (ITL) and puboischiofemoralis (PIF) muscles of male Red Cornish x New Hampshire (RN, 80 d, body weight 2.9 kg) and normal (3.1 kg) broilers were evaluated. 2. In PT muscle the total amount of collagen was significantly greater in RN broilers (3.33 mg/g) than in normal ones (1.71 mg/g). This higher collagen content in RN broilers was based mainly on the closer mesh sizes of endomysial honeycomb. The collagen structures in the perimysia also differed between broiler types, when more collagen fibres were observed in RN broilers. 3. ITL muscle contained total collagen of 4.10 to 5.00 mg/g. Types I and III collagens were distributed on the perimysia at higher percentages in RN broilers (31.6%, 37.2%) than normal (15.6%, 30.8%), respectively. The thick bands of tough collagen fibres characteristic of ITL muscle perimysium in cockerels had not yet developed in these broilers. 4. Total collagen was 4.63 to 6.29 mg/g in PIF material with fascia. In PIF muscle the perimysial collagen fibres had not yet attained their full growth but consisted of densely packed fibrils. PIF muscle was characterised by the earlier maturing collagen structure. 5. These results show that a perimysial collagen structure in broilers is still in an undeveloped state. It is supposed that tenderness of broiler meat is attributed mainly to characteristics of the collagen distribution, in which the majority of types I and III collagens is distributed on the closer mesh of endomysial honeycomb.

Growth changes of the collagen content and architecture in the pectoralis and iliotibialis lateralis muscles of cockerels.

1. Growth changes of the collagen content and architecture in the pectoralis (PT) and iliotibialis lateralis (ITL) muscles were examined using cockerels from 1 to 14 weeks of age. 2. Total collagen content in PT muscle showed little change, but in ITL muscle reached a maximum at 5 weeks and thereafter decreased slightly until 14 weeks. The collagen content was markedly larger in ITL muscle after 5 weeks. Pyridinoline content of collagen increased abruptly from 5 to 14 weeks in both muscles, but no difference between muscle types was detected. 3. The cell size of the endomysial honeycombs increased with the development of myofibres, and the mesh size of the perimysium around the honeycombs enlarged. 4. In both muscles endomysia were an incomplete network of collagen fibrils with many foramina at one week, became a very thin membrane of felt-like fabric in 2 to 5 weeks and thereafter increased in thickness until 11 to 14 weeks. 5. Perimysial width around the secondary fasciculus differed between the muscle types after 5 weeks. In the wider perimysium of ITL muscle, the collagen fibres increased in number and size to make a stack of collagen bands around the fasciculus. In the narrower perimysium of PT muscle, a few platelets of collagen fibres also developed. 6. The perimysial collagen fibre at 1 to 2 weeks had a smooth surface and appeared to be composed of fine collagen fibrils. The fibre at 11 to 14 weeks showed a rugged surface and was composed of coarser collagen bundles that combined with each other into a net-like configuration with very slim meshes. 7. Our results showed that the collagenous components of chicken intramuscular connective tissue changed markedly during the early period of muscle growth in distribution, architecture and quality but with little difference in quantity.

Centrosome hyperamplification and chromosomal damage after exposure to radiation.

OBJECTIVE: In order to elucidate the effects of radiation on centrosome hyperamplification (CH), we examined the centrosome duplication cycle in KK47 bladder cancer cells following irradiation. METHODS: KK47 cells were irradiated with various doses of radiation and were examined for CH immunostaining for gamma-tubulin. RESULTS: Nearly all control cells contained one or two centrosomes, and mitotic cells displayed typical bipolar spindles. The centrosome replication cycle is well regulated in KK47. Twenty-four hours after 5-Gy irradiation, approximately 80% of irradiated cells were arrested in G2 phase, and at 48 h after irradiation, 56.9% of cells contained more than two centrosomes. Laser scanning cytometry performed 48 h after irradiation showed the following two pathways: (1) unequal distribution of chromosomes to daughter cells, or (2) failure to undergo cytokinesis, resulting in polyploidy. With mitotic collection, M-phase cells with CH could be divided into G1 cells with micronuclei and polyploidal cells. Fluorescence in situ hybridization analysis showed clear signs of chromosomal instability (CIN) at 48 h after irradiation. The present study had two major findings: (1) continual duplication of centrosomes occurred in the cell cycle-arrested cells upon irradiation, leading to centrosome amplification; (2) cytokinesis failure was due to aberrant mitotic spindle formation caused by the presence of amplified centrosomes. Abnormal mitosis with amplified centrosomes was detected in the accumulating G2/M population after irradiation, showing that this amplification of centrosomes was not caused by failure to undergo cytokinesis, but rather that abnormal mitosis resulting from amplification of centrosomes leads to cytokinesis block. CONCLUSION: These results suggest that CH is a critical event leading to CIN following exposure to radiation.

Muscle endurance and power spectrum of the triceps brachii in wheelchair marathon racers with paraplegia.

OBJECTIVE: To investigate differences in triceps brachii endurance and electrical activity between elite marathon and recreational wheelchair athletes with paraplegia. DESIGN AND METHODS: Nine male elite wheelchair athletes between 20 and 46 years of age (average 29.0 +/- 8.2 years) with complete (ASIA-A) T4-L1 paraplegia were compared to a group of six male height- and weight-matched recreational wheelchair athletes with similar ages and injuries. Right triceps brachii maximum voluntary contraction (MVC), as well as the duration of the ability to maintain a 50% MVC were determined for all subjects. Median frequency (MF) and mean power frequency (MPF) were evaluated with fast Fourier transform spectrum analysis. MF and MPF rates of change were calculated and compared with the Student's t-test. SETTING: : Department of Rehabilitation Medicine, University of Occupational and Environmental Health, Japan. RESULTS: Right triceps brachii MVCs of the marathoners (42.4 +/- 8.8 N m (range 33-55 N m)) and recreational athletes (41.6 +/- 9.3 N m (range 32-56 N m)) did not differ significantly (P = 0.63). Endurance, however, did. All of the athletes, but none of the control subjects, were able to maintain a 50% MVC contraction of the right triceps brachii for 2 min (the average contraction duration in the latter group was 75.5+/-16.2 s). MF and MPF of the triceps brachii decreased linearly in both groups, but the slopes in the marathoners (-8.9 +/- 4.6 (-3.8 to -16.4) and -9.7 +/- 4.6 (-4.0 to -17.2)%/min, respectively) were statistically less steep than those in the recreational athletes (-22.3 +/- 8.2 (-9.6 to -31.4) and -21.2 +/- 6.4 (-11.4 to -28.6)%/min, respectively). CONCLUSION: Elite marathoners and active wheelchair users have similar triceps brachii strength. The marathoner's triceps brachii, however, display a significantly improved endurance and a slower decline of MF and MPF with time than do those of their recreational athlete control group.

Centrosome hyperamplification and chromosomal instability in bladder cancer.

OBJECTIVE: Chromosomal instability (CIN) is a common feature of malignant tumors. Centrosome hyperamplification (CH) occurs frequently in human cancers, and may be a contributing factor in CIN. In this study, we investigated the relationship between CH and CIN in bladder cancer. METHODS: Clinical samples obtained by transurethral resection from 22 patients with bladder cancer were examined (histological grade G1, 5 cases; G2, 6 cases; G3, 11 cases). CH was evaluated by immunohistochemistry using anti-pericentrin antibody. CIN was evaluated by fluorescence in situ hybridization (FISH). FISH probes for pericentromeric regions of chromosomes 3, 7, and 17 were hybridized to touch preparations of nuclei from frozen tissues. We also analyzed the centrosome replication cycle of bladder cancer by laser scanning cytometry (LSC). RESULTS: Of the 22 cases examined, 18 (81.8%) had centrosome hyperamplification: CH 0, 4 cases (18.1%); CH I, 5 cases (22.7%); CH II, 5 cases (22.7%); CH III, 8 cases (36.4%). The grade of CH was directly proportional to the histological grade (p=0.03, chi(2) test). LSC analysis showed that the centrosome replication cycle was well regulated in pathologically low-grade bladder cancer, which did not have chromosomal instability. In contrast, we found marked variability of centrosomes in pathologically high-grade bladder cancer, which had chromosomal instability. CH and CIN were both detected in pathologically high-grade tumors. The grade of CH was directly proportional to the CIN grade (p=0.0079, chi(2) test). CONCLUSION: The results of the present study suggest that CH may be involved in CIN in bladder cancer.

Relationship among collagen amount, distribution and architecture in the M. longissimus thoracis and M. pectoralis profundus from pigs.

The relative distribution of types I and III collagens and collagen fibre architecture in the perimysium and endomysium were compared to the longissimus thoracis (LT) and pectoralis profundus (PP) muscles in pigs. The LT muscle was composed of type I myofibres 16.8%, IIA 12.9% and IIB 70.2%, and the PP muscle was 25.4, 23.1 and 51.5%, respectively. The total collagen amount differed significantly between the LT (2.66 mg/g) and PP (4.13 mg/g) muscle (P<0.001). On image analysis of the immunohistochemical preparations for types I and III collagens, the percentage area of the perimysium to the total collagen area showed significant differences between the muscles, where perimysial type I collagen occupied 25.4% of the total area in the LT muscle and 45.7% in the PP and perimysial type III 37.6 and 54.5%, respectively (P<0.001). In scanning electron microscopic photographs of the macerated preparation, very thick collagen layers composed of several fibre bands were observed in the perimysia of the PP muscle and appeared differently from the thinner perimysia with a few bands in the LT. Similar architecture of endomysial collagen fibres were observed around every myofibre type in the PP muscle and also in the LT. The fine surface cover of reticular collagen fibres around an adipocyte was shown as a global cast. These results suggested that the total collagen amount of the PP muscle was related mainly to the well developed perimysia composed of several collagen bands, indicating tougher meat compared with the LT muscle.

Biomechanical evaluation of foot pressure and loading force during gait in rheumatoid arthritic patients with and without foot orthosis.

Foot orthoses are commonly used in patients with rheumatoid arthritis (RA) to support the foot and relieve pain, however little is known about the biomechanical effects of in-shoe foot orthoses in reducing or redistributing high pressures and loading forces. The purpose of this study was to compare the foot pressures and loading forces during gait in rheumatoid arthritic patients and healthy subjects, and evaluate the biomechanical effects of the foot orthoses in the RA patients. Twelve female RA patients with foot pain in walking, all Steinbrocker class II, and 8 healthy women without foot pain were matched for age. Foot pressures and loading forces with and without orthoses were measured using the F-Scan program. The pressure distributions and loading forces were standardized by the body weight and compared, and the effects of the foot orthoses were evaluated. The foot orthoses of RA patients provided higher pressure reduction than those of the control group (3.00 +/- 0.38 g/cm2/BW and 3.29 +/- 0.29 g/cm2/BW respectively, p < 0.001). Similar redistribution of plantar pressures and loading forces were found between two groups but the RA patients had a greater change at the stance phase of gait (p < 0.0001). The foot orthosis produces greater pressure and loading force relief and redistribution in RA patients than in normal subjects.