Development of digastric muscles in human foetuses: a review and findings in the flexor digitorum superficialis muscle.

The digastricus and omohyoideus muscles are digastric muscles with two muscle bellies. An insertion tendon of the posterior belly becomes an intermediate tendon in digastricus muscles, whereas a single band-like muscle in omohyoideus muscles may later be interrupted by an intermediate tendon, possibly due to muscle cell death caused by mechanical stress. In human foetuses, an intermediate tendon provides the temporal origins of the tensor veli palatini and tensor tympani muscles. Some reptiles, including snakes, carry multiple series of digastric-like axial muscles, in which each intersegmental septum is likely to become an intermediate tendon. These findings indicate that many pathways are involved in the development of digastric muscles. A review of these morphologies suggested that the flexor digi-torum superficialis (FDS) muscle was a digastric muscle, although the intermediate tendon may not be visible in the surface view in adults. The present observations support the hypothesis that the proximal anlage at the elbow develops into a deep muscle slip to a limited finger, while the distal anlage at the wrist develops into the other slips. The findings suggest that, in the FDS muscle, the proximal and distal bellies of the embryonic digastric muscle fuse together to form a laminar structure, in which muscle slips accumulate from the palmar to the deep side of the forearm. (Folia Morphol 2018; 77, 2: 362-370).

Tensor fasciae latae muscle in human embryos and foetuses with special reference to its contribution to the development of the iliotibial tract.

BACKGROUND: The human tensor fasciae latae muscle (TFL) is inserted into the iliotibial tract and plays a critical role in lateral stabilisation of the hip joint. We previously described a candidate of the initial iliotibial tract that originated from the gluteus maximus muscle and extended distally. MATERIALS AND METHODS: This study extended our observations by examining 30 human embryos and foetuses of gestational age (GA) 7-14 weeks (crown-to-rump length 24-108 mm). At GA 7 weeks, the TFL appeared as a small muscle mass floating in the subcutaneous tissue near the origins of the gluteus medius and rectus femoris muscles. RESULTS: Subsequently, the TFL obtained an iliac origin adjacent to the rectus femoris tendon, but the distal end remained a tiny fibrous mass on the vastus lateralis muscle. Until GA 10 weeks, the TFL muscle fibres were inserted into a vastus lateralis fascia that joined the quadriceps tendon distally. The next stage consisted of the TFL muscle belly "connecting" the vastus fascia and the gluteus fascia, including our previous candidate of the initial iliotibial tract. Until GA 14 weeks, the TFL was sandwiched by two laminae of the connecting fascia. CONCLUSIONS: These findings suggested that, when the vastus lateralis fascia separated from the quadriceps tendon to attach to the tibia, possibly after birth, the resulting iliotibial tract would consist of a continuous longitudinal band from the gluteus maximus fascia, via the vastus fascia, to the tibia. Although it is a small muscle, the foetal TFL plays a critical role in the development of the iliotibial tract.

Foetal development of the human gluteus maximus muscle with special reference to its fascial insertion.

The human gluteus maximus muscle (GMX) is characterised by its insertion to the iliotibial tract (a lateral thick fascia of the thigh beneath the fascia lata), which plays a critical role in lateral stabilisation of the hip joint during walking. In contrast, in non-human primates, the GMX and biceps femoris muscle provide a flexor complex. According to our observations of 15 human embryos and 11 foetuses at 7-10 weeks of gestation (21-55 mm), the GMX anlage was divided into 1) a superior part that developed earlier and 2) a small inferior part that developed later. The latter was adjacent to, or even continuous with, the biceps femoris. At 8 weeks, both parts inserted into the femur, possibly the future gluteal tuberosity. However, depending on traction by the developing inferior part as well as pressure from the developing major trochanter of the femur, most of the original femoral insertion of the GMX appeared to be detached from the femur. Therefore, at 9-10 weeks, the GMX had a digastric muscle-like appearance with an intermediate band connecting the major superior part to the small inferior mass. This band, most likely corresponding to the initial iliotibial tract, extended laterally and distally far from the muscle fibres. The fascia lata was still thin and the tensor fasciae latae seemed to develop much later. It seems likely that the evolutionary transition from quadripedality to bipedality and a permanently upright posture would require the development of a new GMX complex with the iliotibial tract that differs from that in non-human primates. (Folia Morphol 2018; 77, 1: 144-150).

A new insight into the fabella at knee: the foetal development and evolution.

Using longitudinal semiserial sections of 12 lower extremities from 8 human foetuses at 15-18 weeks, we compared foetal morphologies of the knee in specimens with and without fabellae. We also compared the fabella, if present, with the hallucal sesamoid in the same foetus. Cartilaginous fabella, positive for versican and tenascin by immunohistochemistry, was found in 5 of the 8 foetuses. This structure was embedded in a thick and tight lateral fibrous band, providing a common origin of the plantaris muscle and the lateral head of the gastrocnemius muscle. The plantaris was covered by the lateral head of the gastrocnemius, but these 2 muscles were separated by a distinct fascia or space. Notably, the foetal fabella did not attach to the joint capsule. In the 3 specimens without fabellae, the lateral fibrous band was thin, containing a fibrous mass, negative for versican and tenascin, in place of the fabella. The "medial" head of the gastrocnemius faced or covered the plantaris, while the lateral head was continuous with the plantaris. A hallucal cartilaginous sesamoid, positive for versican and tenascin, was present in all 8 specimens. It carried a flat surface facing the joint cavity and was covered by tendons of the short muscles of the foot. Because of the difference in topographical relation of muscles between specimens with or without fabella, rather than mechanical stress to the tendon, fabella development may require a distinct plantaris muscle independent of the gastrocnemius. We discussed about an evolutionary aspect of the fabella and plantaris muscle.

Oblique cord (chorda obliqua) of the forearm and muscle-associated fibrous tissues at and around the elbow joint: a study of human foetal specimens.

In adults, the oblique cord or chorda obliqua separates the origins of the flexor pollicis longus (FPL) and flexor digitorum profundus (FDP) muscles from the supinator muscle and elbow joint. This study examined the topographic anatomy of the oblique cord and related muscles in foetuses. Semiserial sections of five mid-term foetuses of gestational age (GA) 14-16 weeks and 12 late-stage foetuses of GA 28-30 weeks were histologically examined and three forearms at GA 30 weeks were macroscopically evaluated. Late-stage foetuses showed a fascial structure between the supinator and FDP muscles. The latter extended proximally to the elbow joint and the muscle origin thickened the distal, ulnar part of the capsule. The FPL origin also extended proximally but did not reach the joint capsule. These morphologies were consistent with macroscopic examinations. The brachialis muscle was widely inserted into the proximal, anterior part of the capsule. In addition, the medial collateral ligament was not covered by the pronator-flexor muscles but by the triceps brachii muscle. The oblique cord apparently did not form prenatally. After birth, the proximal parts of the FDP and FPL muscles were likely replaced by collagenous tissues, providing a specific type of intermuscular septum i.e., the oblique cord. This type of muscle-ligament transition was observed in the annular ligament of the radius. The foetal elbow joint was characterised by strong support by the FDP, brachialis and triceps brachii muscles. Therefore, the foetal elbow is not a miniature version of the adult elbow.

Positional changes in tendon insertions from bone to fascia: development of the pes anserinus and semimembranosus muscle insertion in human foetuses.

Development of a long muscle belly in foetal extremities generally requires a definite bony insertion of the long tendon. However, in adults, the pes anserinus and the semimembranosus tendon (SMT) are inserted into fasciae. Development of fascial insertions in foetuses was investigated by examining serial histological sections obtained from 7 foetuses at 8-9 weeks and 8 foetuses at 14-16 weeks. The presence of matrix substances and macrophages was also examined by immunohistochemistry. At 8 weeks, the tendons of the semitendinosus, gracilis, sartorius and semimembranosus muscles were straight and inserted into the initial shaft-like proximal end of the tibia on the proximal side of the popliteus muscle. At 9 weeks, however, the medially extending popliteus muscle appeared to push the pes anserinus tendons superficially, with a loss of cartilage insertions. The SMT obtained an attachment to the popliteus muscle. At 14-16 weeks, the SMT divided into thick and thin bundles: the former contained abundant macrophages and inserted into the tenascin-positive perichondrium of the enlarged proximal tibia, while the later without macrophages ended at the joint capsule. The pes anserinus tendons, negative for both versican and tenascin-c, took highly tortuous courses toward the fascia cruris. Because the medial extension of the popliteus muscle was associated with the enlargement of the proximal tibia, the topographical relationship of the popliteus muscle with these 4 tendons changed drastically, resulting in a loss of cartilage insertion of the pes anserinus tendons as well as the division and reconstruction of the SMT.

Lymphatic vessels around the uterus: D2-40 (podoplandin) immunohistochemistry using elderly cadavers.

Using D2-40 immunohistochemistry, we examined the morphology of lymphatic vessels (LVs) in, along and around the uterus of 10 donated female cadavers (mean age, 85 years). All these women had 1 or 2 children with vaginal delivery, but the other obstetrics information was unknown. When compared with the bladder, vagina and the subperitoneal tissue, the percentage area of LVs in a 3 × 2 mm square including the hot spot was extremely high along the uterine artery and superficial uterine vein, in spite of the silent physiology of the elderly uterus. Notably, the LVs along the uterine artery and superficial uterine vein were highly dilated and embedded in the tight connective tissue around the artery and vein. In contrast, the LVs were separated from the artery and vein in the so-called vesico- -uterine ligament. Thus, surgical separation of the LVs from the artery and vein, i.e., skeletonisation, appears very difficult along the uterine artery and superficial uterine vein. This may become a major factor limiting the future application of robot-assisted surgery for uterine cancers.

Cavitation processes in a space filled with loose mesenchymal tissues: a comparison between the retrosternal space and the middle ear tympanic cavity in human fetuses.

BACKGROUND: During the expansion of the pleural cavity in early fetuses, a thick sheet of loose mesenchymal tissue (SLMT) appears between the lung bud and body wall. Subsequently, the growing lung bud invades into the SLMT and the latter becomes fragmented to disappear. To compare this with the tympanic cavity filled with loose mesenchymal tissues, the present study aimed to demonstrate the development, establishment, and breaking of the SLMT in the retrosternal space. MATERIALS AND METHODS: Although the retrosternal tissue was almost absent or very thin at 7 weeks, the SLMT appeared behind the manubrium sterni at 8 weeks. Accordingly, at 9-10 weeks, cavitation occurred in the SLMT to expand the pleural cavity. Therefore, the volume of SLMT was not determined by the adjacent structures such as the pericardium and sternum. Likewise, mesenchymal tissues filling the middle ear disappeared after 26 weeks. RESULTS: There were considerable individual variations in the timing of beginning and location of the tympanic cavitation. However, in contrast to the retrosternal SLMT, the volume of the future tympanic cavity is determined by the adjacent hard tissue and tympanic membrane much earlier than the cavitation. The mesenchymal tissue carried abundant vessels in the middle ear but none or few veins in the retrosternal SLMT. CONCLUSIONS: The concept that the lung bud invades into the splanchnic mesoderm to expand the pleural cavity seems oversimplified. Mechanical stresses from the pleural cavity might induce retrosternal cavitation, while a loss in blood supply might cause tympanic cavitation.

Inferior oblique muscle of the eye: its foetal development with special reference to understanding of the frequent variants in adults.

BACKGROUND AND MATERIALS AND METHODS: To provide better understanding of frequent variations of the inferior oblique (IO) of adult extraocular muscles, we observed sagittal and horizontal histological sections of the eye and orbits from 32 foetuses (7-34 weeks of gestational age; 24-295 mm of crown-rump length). RESULTS: In early foetuses (7-8 weeks), the IO was restricted at an antero-infero-medial angle of the future orbit. In contrast to extraocular recti, the IO appeared to extend along the mediolateral axis and had no definite tendon. At midterm, the IO tendon became evident. Sometimes, the IO muscle belly attached to the inferior rectus or, the IO tendon divided into two laminae to enclose the lateral rectus. At late-term, a multilayered sheath was evident around the sclera and, via one or some of the fascial layers, the IO was communicated with a fascia enclosing the inferior rectus. At midterm and late-term, the IO originated not only from the maxilla near the orbicularis oculi origin but also from a vein-rich fibrous tissue around the lacrimal sac. Both origins were muscular without intermittent tendon or ligament. Therefore, the fascial connection as well as a direct contact between the IO and the inferior or lateral rectus seemed to provide variant muscular bridges as reported in adults. Moreover, the two attachment sites at the origin seemed to provide double muscle bellies of the adult IO. CONCLUSIONS: Consequently, the present specimens contained seeds of any types of adult variations. The muscle fibres from the lacrimal sac might play a role for the lacrimal drainage.

Development and growth of the foot lumbricalis muscle: a histological study using human foetuses.

BACKGROUND: Our group has shown early development of the hand lumbricalis and hypothesized that, at midterm, the lumbricalis (LU) bundles flexor tendons to provide a configuration of "one tendon per one finger" (Cho K.H. Folia Morphol. 2012; 71, 3: 154-163). However, the study concentrated on the hand and contained no sections of near-term foetuses. MATERIALS AND METHODS: The present examination of paraffin-embedded tangential sections along the planta from 25 embryos and foetuses at 6-40 weeks (15-320 mm crown-rump length) demonstrated that, at 8 weeks, the initial foot LU appeared in the proximal side of the common tendinous plate of all five deep tendons. RESULTS: After midterm, a drastic three-phase change occurred at the muscle origin: 1) the LU originated from each of the flexor digitorum longus tendon (FDLT), but abundant tenocyte candidates separated the muscle fibre from the tendon collagen bundle; 2) the LU arose from the covering fascia depending on increased thickness of the muscle; and 3) the LU muscle fibres intermingled with tendon collagen bundles and partly surrounded the tendon. Simultaneously, a dividing site of the FDLT migrated distally to accelerate the changes at the LU origin. These phases did not always correspond to the size of foetus after 30 weeks. CONCLUSIONS: Consequently, in contrast to the hand LU, the delayed changes in the foot were characterised by involvement of the LU origin into a single common part of the FDLT. The quadratus plantae muscle fibres did not attach to the LU at any phase, and connected with the fourth and fifth toe tendons.

Changes in topographical relation between the ductus arteriosus and left subclavian artery in human embryos: a study using serial sagittal sections.

BACKGROUND: At birth, the ductus arteriosus (DA) merges with the aortic arch in the slightly caudal side of the origin of the left subclavian artery (SCA). Since the SCAs (7th segmental arteries) were fixed on the level of the 7th cervical-first thoracic vertebral bodies, the confluence of DA should migrate caudally. We aimed to describe timing and sequence of the topographical change using serial sagittal sections of 36 human embryos and foetuses (CRL 8-64 mm; 5-10 weeks), Those made easy evaluation of the vertebral levels possible in a few section. MATERIALS AND METHODS: The DA or 6th pharyngeal arch artery seemed to slide down in front of the sympathetic nerve trunk along 1.0-1.2 mm from the second cervical vertebral level at 5-6 weeks and, at 6 weeks (CRL 14-17 mm), the DA confluence with aorta reached the 7th cervical level. Because of the highly elongated common carotid artery, the sliding of DA confluence seemed to be much shorter than the cervical vertebrae growing from 1 mm to 2.4 mm. RESULTS: At the final topographical change at 6-7 weeks, the DA confluence further descended to a site 1-vertebral length below the left SCA origin. From 6 to 9 weeks, a distance from the top of the aortic arch to the left SCA origin was almost stable: 0.3-0.5 mm at 6 weeks and 0.4-0.6 mm at 9 weeks. CONCLUSIONS: The heart descent and the caudal extension of the trachea and bronchi, those occurred before the DA sliding, were likely to be a major driving force for the sliding.

Observations of foetal heart veins draining directly into the left and right atria.

Evaluation of semiserial sections of 14 normal hearts from human foetuses of gestational age 25-33 weeks showed that all of these hearts contained thin veins draining directly into the atria (maximum, 10 veins per heart). Of the 75 veins in these 14 hearts, 55 emptied into the right atrium and 20 into the left atrium. These veins were not accompanied by nerves, in contrast to tributaries of the great cardiac vein, and were negative for both smooth muscle actin (SMA) and CD34. However, the epithelium and venous wall of the anterior cardiac vein, the thickest of the direct draining veins, were strongly positive for SMA and CD34, respectively. In general, developing fibres in the vascular wall were positive for CD34, while the endothelium of the arteries and veins was strongly positive for the present DAKO antibody of SMA. The small cardiac vein, a thin but permanent tributary of the terminal portion of the great cardiac vein, was also positive for SMA and CD34. A few S100 protein-positive nerves were observed along both the anterior and small cardiac veins, but no nerves accompanied the direct dra- inage veins. These findings suggested that the latter did not develop from the early epicardiac vascular plexus but from a gulfing of the intratrabecular space or sinus of the atria. However, the immunoreactivity of the anterior cardiac vein suggests that it originated from the vascular plexus, similar to tributaries of the great cardiac vein.

Differences in foetal topographical anatomy between insertion sites of the iliopsoas and gluteus medius muscles into the proximal femur: a consideration of femoral torsion.

BACKGROUND: Prenatal twisting of the femoral neck seems to result in an angle of anteversion or torsion, but the underlying process has not been elucidated. MATERIALS AND METHODS: This study analysed sagittal, frontal and horizontal sections of 34 embryo and foetal specimens of gestational age (GA) 6-16 weeks (crown-rump length 21-130 mm). At GA 6-7 weeks, the iliopsoas (IP) and gluteus medius (GME) muscles were inserted into the anterior and posterior aspects of the femur, respectively, allowing both insertions to be viewed in a single sagittal section. RESULTS: At GA 8 weeks, the greater trochanter and the femoral neck angle became evident, and the GME tendon was inserted into the upper tip of the trochanter. At GA 9 weeks, the location of IP insertion was to the medial side of the GME insertion. After 9 weeks, the IP insertion consisted of a wavy, tendino- us part of the psoas muscle and another part of the iliacus muscle, with many fibres of the latter muscle attached to the joint capsule. After GA 12 weeks, the IP was inserted into the anteromedial side of the greater trochanter, while the aponeurotic insertion of the GME wrapped around the trochanter. At GA 15-16 weeks, a deep flexion at the hip joint caused an alteration in the relative heights of the lesser and greater trochanter, with the former migrating from the inferior to the slightly superior side. CONCLUSIONS: These findings indicate that twisting of the femoral neck started at GA 8-9 weeks.

Glutamine preferentially inhibits T-helper type 2 cell-mediated airway inflammation and late airway hyperresponsiveness through the inhibition of cytosolic phospholipase A(2) activity in a murine asthma model.

BACKGROUND: The non-essential amino acid, l-glutamine (Gln), is abundant in the human body. Gln exhibits beneficial effects on endotoxic shock through the inhibition of cytosolic phospholipase A(2) (cPLA(2)) activity. cPLA(2) has been reported to be implicated in the pathogenesis of asthma, but the effects of Gln on asthma have not yet been defined. OBJECTIVE: To investigate the effects of Gln on allergic bronchial inflammation and airway hyperresponsiveness (AHR), and to determine the possible action mechanisms of Gln in a murine model of asthma. METHODS: cPLA(2) phosphorylation was assessed by immunoprecipitation and Western blotting. Smears of bronchoalveolar lavage cells were stained with Diff-Quik solution for differential cell counting. Airway levels of the proteins [T-helper type-1 (Th1) and Th2 cytokines, and mucin] were measured by ELISA. mRNA expression of cytokines was assessed by real-time RT-PCR. AHR was assessed as a change in airway resistance (RL). Histological studies were performed to assess the levels of mucin and pulmonary inflammation. RESULTS: Systemic Gln administration inhibited cPLA(2) phosphorylation and its enzymatic activity in the lungs. Additionally, Gln effectively suppressed the key features of Th2-dependent asthmatic features, such as airway eosinophilia, mucus formation, and airway type 2 cytokine production, as well as late AHR. CONCLUSION: Gln was found to be effective in the suppression of Th2-dependent phenotypes and late AHR, and this effect of Gln appeared to be at least partially attributable to its ability to suppress cLPA(2) activity in the airway. Our results suggest that clinical use of Gln for patients with asthma may be beneficial.

Isolation of a strain of Hantaan virus from peritoneal exudate cells of a patient with hemorrhagic fever with renal syndrome.

A new strain of Hantaan virus (HTNV), GH716, was isolated from the peritoneal exudate cells (PEC) of a patient with severe hemorrhagic fever with renal syndrome (HFRS). The isolate was propagated in Vero E6 cells. At each passage the virus-infected cells were examined for HTNV with immunofluorescence technique using monoclonal antibodies (McAb) 25-1 and 84-1 against HTNV. From passage 5 on, fluorescent intensity tended to stabilize at (+++) and infectivity titer reached 10(5) TCID50/ml. Using 14 McAb to 76-118 and BI strains of HTNV, we found that strain GH716 was antigenically similar to strain 76-118 (Apodemus type) and different from strain R22 (Rattus type), suggesting that GH716 may fall into the Apodemus type of HTNV. This is the first isolation of an HTNV from human PEC collected on the tenth day of illness. The successful isolation of strain GH716 may provide an alternative source of obtaining HTNV during the later stages of HFRS.

Immunodeficiency in patients with hemophilia: an underlying deficiency and lack of correlation with factor replacement therapy or exposure to human immunodeficiency virus.

Nineteen patients with hemophilia, five of whom were classified as untreated, were immunologically evaluated by the measurement of cell surface markers, natural killer cell activity, mitogenic responses, polyclonal immunoglobulin production, and serologic evidence of human immunodeficiency virus infection. No correlations were found between human immunodeficiency virus infection and immune abnormalities, or between patients who did or did not receive factor therapy, although the abnormalities were more profound in those who received treatment. An intrinsic B cell abnormality in patients with hemophilia is suggested.

Inhibition of suppressor cell function by cimetidine in a murine model.

Immunomodulatory effects of cimetidine, an H2 histamine receptor antagonist, have been reported in humans and animals. To define these effects more clearly, the action of cimetidine on suppressor cell function was studied utilizing a murine model of contact hypersensitivity. Intravenous inoculation of BALB/c mice with DNP-coupled syngeneic spleen cells induced the production of DNP-specific suppressor cells which could easily be demonstrated by a reduction in ear swelling after contact sensitization with 1-fluoro-2,4-dinitrobenzene (DNFB) following transfer of spleen and lymph node cells to naive syngeneic recipients. Cimetidine treatment of animals in which suppressor cells were induced resulted in an inability of these mice to transfer cellular suppression as measured by development of a normal immunologic response in the recipient mice. The effect of cimetidine was both dose and time related. While all groups receiving cimetidine showed some loss of suppressor cell function, the maximum effect (up to 100% inhibition) was seen when 50 mg/kg of cimetidine was administered intraperitoneally 2 days before or on the day of suppressor cell induction. Some restoration also occurred when cimetidine was given after the day of induction. It has been shown that suppressor cells possess histamine receptors which may be involved in suppressor cell activation. The results indicate that cimetidine may inhibit the functioning of these receptors.