The role of endocytosis in regulating L1-mediated adhesion

L1 is a neural cell adhesion molecule critical for neural development. Full-length L1 (L1(FL)) contains an alternatively spliced cytoplasmic sequence, RSLE, which is absent in L1 expressed in nonneuronal cells. The RSLE sequence follows a tyrosine, creating an endocytic motif that allows rapid internalization via clathrin-mediated endocytosis. We hypothesized that L1(FL) would internalize more rapidly than L1 lacking the RSLE sequence (L1(Delta)(RSLE)) and that internalization might regulate L1-mediated adhesion. L1 internalization was measured by immunofluorescence microscopy and by uptake of (125)I-anti-rat-L1 antibody, demonstrating that L1(FL) is internalized 2-3 times faster than L1(Delta)(RSLE). Inhibition of clathrin-mediated endocytosis slowed internalization of L1(FL) but did not affect initial uptake of L1(Delta)(RSLE). To test whether L1 endocytosis regulates L1 adhesion, cell aggregation rates were tested. L1(Delta)(RSLE) cells aggregated two times faster than L1(FL) cells. Inhibition of clathrin-mediated endocytosis increases the aggregation rate of the L1(FL) cells to that of L1(Delta)(RSLE) cells. Our results demonstrate that rapid internalization of L1 dramatically affects L1 adhesion.     

Effects of losartan, in monotherapy or in association with hydrochlorothiazide, in chronic nephropathy resulting from losartan treatment during lactation

We recently standardized a model (L(Lact)) of severe chronic kidney disease based on impaired nephrogenesis by suppression of angiotensin II activity during lactation (Machado FG, Poppi EP, Fanelli C, Malheiros DM, Zatz R, Fujihara CK. Am J Physiol Renal Physiol 294: F1345-F1353, 2008). In this new study of the L(Lact) model, we sought to gain further insight into renal injury mechanisms associated with this model and to verify whether the renoprotection obtained with the association of the angiotensin II receptor blocker losartan (L) and hydrochlorothiazide (H), which arrested renal injury in the remnant kidney model, would provide similar renoprotection. Twenty Munich-Wistar dams, each nursing six pups, were divided into control, untreated, and L(Lact) groups, given losartan (L; 250 mg·kg(-1)·day(-1)) until weaning. The male L(Lact) offspring remained untreated until 7 mo of age, when renal functional and structural parameters were studied in 17 of them, used as pretreatment control (L(Lact)Pre), and followed no further. The remaining rats were then divided among groups L(Lact)+V, untreated; L(Lact)+L, given L (50 mg·kg(-1)·day(-1)) now as a therapy; L(Lact)+H, given H (6 mg·kg(-1)·day(-1)); and L(Lact)+LH, given L and H. All parameters were reassessed 3 mo later in these groups and in age-matched controls. At this time, L(Lact) rats exhibited hypertension, severe albuminuria, glomerular damage, marked interstitial expansion/inflammation, enhanced cell proliferation, myofibroblast infiltration, and creatinine retention. L monotherapy normalized albuminuria and prevented hypertension and the progression of renal injury, inflammation, and myofibroblast infiltration. In contrast to the remnant model, the LH combination promoted only slight additional renoprotection, perhaps because of a limited tendency to retain sodium in L(Lact) rats.     

Rhythmic Conidiation in Constant Light in Vivid Mutants of Neurospora crassa

In Neurospora crassa, a circadian rhythm of conidiation (asexual spore formation) can be seen on the surface of agar media. This rhythm has a period of 22 hr in constant darkness (D/D). Under constant illumination (L/L), no rhythm is visible and cultures show constant conidiation. However, here we report that strains with a mutation in the vivid (vvd) gene, previously shown to code for the photoreceptor involved in photo-adaptation, exhibit conidiation rhythms in L/L as well as in D/D. The period of the rhythm of vvd strains ranges between 6 and 21 hr in L/L, depending upon the intensity of the light, the carbon source, and the presence of other mutations. Temperature compensation of the period also depends on light intensity. Dark pulses given in L/L shift the phase of the rhythm. Shifts from L/L to D/D show unexpected after effects; i.e., the short period of a vvd strain in L/L gradually lengthens over 2–3 days in D/D. The rhythm in L/L requires the white collar (wc-1) gene, but not the frequency (frq) gene. FRQ protein shows no rhythm in L/L in a vvd strain. The conidiation rhythm in L/L in vvd is therefore driven by a FRQ-less oscillator (FLO).     

The genus Leishmania in Italy

Seventy-four Leishmania isolates collected in Italy from six different Regions where leishmaniases are endemic, have been typed. Parasites have been isolated from: man (VL and CL), dog, black rat (Rattus rattus), fox (Vulpes vulpes) and geckoes (Tarentola mauritanica and Cyrtodactylus kotschyi). The isolates have been characterized by starch-gel electrophoresis for 9-16 enzymes whose mobility was compared with that of international reference strains for L. infantum, L. tropica, L. major, L. donovani, L. aethiopica and L. tarentolae. The results obtained have shown that the genus Leishmania in Italy is represented by five zymodemes which may be grouped into two taxa: L. infantum s.l. (L. infantum s.st., L. infantum NH130 variant, L. infantum NH140 variant and L. infantum GOT, MDH, NH variant), agent of mammalian leishmaniases (including human leishmaniases), and L. tarentolae, parasite of geckoes. At the moment, the absence of L. tropica in Italy as agent of CL has been revealed. Through the analysis of epidemiological data obtained from the foci where Leishmania parasites were isolated two zymodemes only, L. infantum s.st. and L. infantum NH140 variant, show to be widely distributed. However, L. infantum s.st. appears to be prevalent in Thyrrenean foci which are characterized by VL cases and by high density of Phlebotomus perniciosus, and L. infantum NH140 variant is present in Adriatic areas where CL is diffuse and P. perfiliewi is the probable vector.     

Ezetimibe restores biliary cholesterol excretion in mice expressing Niemann-Pick C1-Like 1 only in liver

Niemann-Pick C1-Like 1 (NPC1L1) is highly expressed in the small intestine across mammalian species and is the target of ezetimibe, a potent cholesterol absorption inhibitor. In humans, NPC1L1 is also expressed in the liver. We found that transgenic overexpression of NPC1L1 in the wild-type mouse liver inhibits biliary cholesterol secretion and raises blood cholesterol, which can be reversed by ezetimibe treatment. Unfortunately, the high expression of endogenous NPC1L1 in the intestine hampered a definitive establishment of the role of hepatic NPC1L1 in cholesterol metabolism and ezetimibe action in the liver because intestinal NPC1L1 dramatically influences cholesterol homeostasis and is a target of ezetimibe. To circumvent this obstacle, we crossed liver-specific NPC1L1 transgenic mice to NPC1L1 knockout (L1-KO) mice and created a mouse line expressing no endogenous NPC1L1, but human NPC1L1 in liver only (L1(LivOnly) mice). Compared to L1-KO mice, L1(LivOnly) mice on a 0.2% cholesterol diet showed significantly increased hepatic and plasma cholesterol, and despite a 90% reduction in biliary cholesterol excretion, their fecal cholesterol excretion remained completely unaltered. Remarkably, 4days of ezetimibe treatment significantly restored biliary cholesterol secretion in L1(LivOnly) mice. These findings demonstrated a direct role of hepatic NPC1L1 in regulating biliary cholesterol excretion and hepatic/blood cholesterol levels, and unequivocally established hepatic NPC1L1 as a target of ezetimibe.     

Plasma chemistry reference values in ostriches

Reference values for 18 plasma chemical variables in blue neck ostriches (Struthio camelus australis, n = 60, age 24-36 mo) were established for use in veterinary clinical practice using nonparametric statistics. The following values were established for the percentiles P2.5 and P97.5: sodium 147-157 mmol/L, calcium 2.4-4.8 mmol/L, inorganic phosphate 1.3-2.3 mmol/L, chloride 94-105 mmol/L, glucose 10.3-13.7 mmol/L, urea 0.5-0.8 mmol/L, uric acid 351-649 mumol/L, bile acids 8-33 mumol/L, total protein 39-56 g/L, albumin-globulin ratio 0.45-0.59, osmolality 304-330 mOsm/kg, alkaline phosphate 69-217 IU/L, aspartate aminotransferase 243-418 IU/L, gamma-glutamyltransferase 0-1 IU/L, creatine kinase 1648-4894 IU/L, glutamate dehydrogenase 8-17 IU/L, and lactate dehydrogenase 860-2236 IU/L. The plasma calcium concentration was significantly (P < 0.001; r = 0.74) related to the total protein concentration and an adjustment-formula for calcium was derived: adjusted Ca (mmol/L) = Ca (mmol/L)--0.09 TP (g/L) + 4.4. The influence of blood sample treatment on the plasma potassium concentration as seen in other avian species was demonstrated in a separate experiment, emphasizing the need to separate plasma and cells immediately after collection in avian blood samples.     

Cell population kinetics in pig epidermis: further studies

The cell population kinetics of the epidermis were studied in 4-month-old pigs. Mitotic figures were confined to the basal cell (L1) and the first suprabasal cell layer (L2). The mitotic index (MI) was 0.17 +/- 0.04% for L1 and 0.08 +/- 0.03% for L2. Labelled nuclei were distributed throughout the viable epidermis, the majority (79.1 +/- 1.1%) were in L1 with 19.5 +/- 1.2% in L2. The labelling indices (LI) in layers L1 and L2 were 7.1 +/- 0.4% and 3.4 +/- 0.1%, respectively. After labelling with two injections of tritiated thymidine [3H]TdR separated by 90 min, the LI increased to 8.2 +/- 0.3% in L1 and to 4.0 +/- 0.2% in L2. This increased labelling confirmed that cell proliferation occurs in both layers, L1 and L2, of the epidermis. The cell production rate (K) in L1 and L2 had an upper limit of 10.7 +/- 1.0 and 6.2 +/- 1.8 cells per 1000 cells per hour respectively. The cell flow rate per hour (cell flux), into and out of the DNA synthesis phase (S), and the duration of DNA synthesis were determined from double-labelling studies with [3H]TdR and [14C]TdR. The cell flux into and out of S was identical and was calculated as 0.6 +/- 0.1%/hr (L1) and 0.5 +/- 0.1%/hr (L2). Values for tS varied from 8 to 10 hr. The cell turnover times (tT) were in the range 89-129 hr and 180-261 hr for L1 and L2, respectively. Log normal curves were fitted to the fraction labelled mitoses data for L1 and L2. Values for tS for cells in L1 and L2 were 9.8 hr and 11.9 hr, respectively. tG2 + 1/2tM was 7.2 hr in L1 and 9.1 hr in L2.     

Methylation of Ribosomal Proteins in Escherichia coli

Escherichia coli was grown in a medium containing [1-(14)C]methionine and [methyl-(3)H]methionine, and the (3)H/(14)C ratio was determined for each of the ribosomal proteins derived from the 70S ribosome. Evidence indicates that six proteins from the 50S subunit were methylated: L7, L9, L11, L12, L18, and L33. Methylation of several other 50S proteins (such as L1, L3, L5, etc.) may also occur. The methylated amino acids in protein L11 have been characterized further and found to be predominately epsilon-trimethyllysine. A small amount of a compound tentatively identified as N(G), N'(G)-dimethylarginine was also detected.     

Further Studies on Leptospirosis in Small Rodents and Shrews in Finland. A Report on Investigations Made in 1963-64

In previous investigations on small mammals in Finland (Rislakki & al 1954, Rislakki & Salminen 1955, Salminen 1956), the Leptospira icterohaemorrhagiae-frequency in rats (Rattus norvegicus) was rather high (43.1 %). Leptospira-positive cases were also found in house mice (L. sejroe 23.3 %), harvest mice (L. hataviae 9.0 %), yellow necked field mice (L. poi 12.5 %), common voles (L. sejroe and L. bataviae together 12.1 %), field voles (L. sejroe and L. bataviae together 10.7 %) and in common shrews (L. poi 1.2 %). Specimens of other species sent in for investigation (Norway rat, common red backed vole, large tooth backed vole, northern red backed vole, root vole, water vole, wood lemming, Laxmann's shrew, lesser shrew and water shrew) gave negative results.     

Phlebotominae sand flies in Paraguay: abundance distribution in the Southeastern region

From September 1993 to August 2001, 7,190 phlebotomine were collected with CDC light trap in an endemic area for human leishmaniasis, in the departments of Misiones and Itap a, Paraguay. Eleven species were identified: Lutzomyia neivai (93.7%), L. whitmani (4.1%), and L. fischeri, L. shannoni, L. migonei, L. misionensis, L. cortelezzii, L. pessoai, L. alphabetica, Brumptomyia avellari and B. guimaraesi (less than 1%). The last three species are new records for the country. The biodiversity and phlebotomine abundance were associated with the proximity to primary forest or gallery forest, but L. neivai was also found in peridomestic periurban environment. L. neivai was found throughout the year, and showed a period of higher activity from September to April (spring to fall) with a unimodal or bimodal pattern in relation to the annual rainy peaks during the summer. Background literature about phlebotomine from Paraguay has been reviewed.     

Blood and urine physiological values in farm-cultured Rana catesbeiana (Anura: Ranidae) in Argentina

A total of 302 samples of healthy farm-cultured Rana catesbeiana specimens (9-21 months-old, 50-350 g liveweight, 50% each sex) from the north-east of Argentina, were analyzed through spectrophotometry, electrophoresis, densitometry, refractometry and microscopy in order to obtain blood and urine normal values. Confidence intervals (p<0.05) for PCV (28.6-31.6%), RBC (0.40-0.44 T/L), MCV (686-732 fL), hemoglobin (6.41-7.20 g/dL), MCH (151-164 pg), MCHC (22.6-24.0%), WBC (18.7-22.3 G/L), neutrophils (58.4-63.4%), lymphocytes (23.9-29.8%), monocytes (2.1-3.8%), eosinophils (4.6-7.0%), basophils (2.9-4.1%), bleeding time (289-393s), coagulation time (452-696s), prothrombin time (76-128s), urinary density (1.0061-1.0089 g/mL), urinary pH (6,38-6.96)., fibrinogen (0.59-0.99 g/dL), total protein (4.19-4.49 g/dL), albumin (1.49-1.67 g/dL), alpha-1 globulin (0.20-0.24 g/dL), alpha-2 globulin (0.48-0.54 g/dL), beta globulin (0.68-0.77 g/dL), gamma globulin (1.28-1.42 g/dL), albumin/globulin ratio (0.50-0.58), creatinine (4.09-5.56 mg/L). urea (76.1-92.4 mg/L), uric acid (11.5-15.4 mg/L), triglycerides (0.34-0.52 g/L), total cholesterol (0.56-0.67 g/L), HDL-C (0.03-0.05 g/L), LDL-C (0.34-0.44 g/L), alpha lipoprotein (6.01-8.67%). beta lipoprotein (91.3-93.9%), glucose (0.45-0.54 g/L), Na (116-121 meq/L), K (3.42-3.81 meq/L), Cl (100-116 meq/L), Ca (7.98-8.61 mg/dL). P (8.319.36 mg/dL), Mg (2.26-2.55 mg/dL), Fe (105-178 ug/dL), ALP (144-170 [U/L), ALT (10.0-14.8 IU/L), AST (42.8-53.4 IU/L), GGT (7.8-10.6 IU/L), LDH (99-135 IU/L), CHE (151-185 lU/L) and CPK (365-500 IU/L), were obtained. Some parameter ranges were similar to those obtained in amphibians, birds or mammals; others were very different. These parameters are useful to evaluate sanitary, metabolic and nutritional state on captive bullfrogs.     

The OKRA leaf shape mutation in cotton is active in all cell layers of the leaf

Okra (L2O) is a semidominant mutation of cotton (Gossypium barbadense) that alters leaf shape by increasing the length of lobes and decreasing lamina expansion. Chimeras containing L2O and wild-type tissue were generated using Semigamy (Se), a mutation that blocks syngamy during fertilization and produces haploid maternal/paternal chimeral progeny at low frequency. In sectorial chimeras, changes in leaf morphology coincide with the boundary between mutant and wild-type tissues, suggesting that L2O does not regulate a laterally diffusible factor within the leaf. However, in mericlinal or periclinal chimeras, the presence of L2O in tissue derived from any of the three histogenic layers (L1, L2, or L3) of the shoot apical meristem produced leaves with a partial mutant phenotype. The presence of L2O in the epidermis (an L1 derivative), or in the subepidermal mesophyll of the leaf (L2 derivatives) reduced the growth of the lamina and thus increased the depth of leaf lobes. The presence of L2O in the middle mesophyll of the lamina and the vasculature of major lateral veins (L3 derivatives) had no local effect on the expansion of the lamina, but significantly increased lobe length. These results demonstrate that L2O is active in every tissue layer of the leaf.     

The reclassification of Lycopodiaceae (s. str.) in China

The fern-allied family Lycopodiaceae(s. str. ) in China is reclassified in the present paper. Six genera, fifteen species and three forms are recorded. The genus Pseudolycopodiella Holub (1983) is adopted, one new name, Lycopodium neopungens H. S. Kung et L. B. Zhang, is given to replace the invalid name L. pungens Desv., one new form is described: Palhinhaea hainanensis C. Y. Yang f. glabra H. S. Kung et L. B. Zhang, and nine names are treated for the first time as synonyms: L. annotinum L. var. brevifolium Christ( = L. zonatum Ching), L. annotinum L. var. aciculare Christ( = L. zonatum Ching), L. alticola Ching( = L. zonatum Ching), L. simulans Ching et H. S. Kung ex Ching( = L. japonicum Thunb. ex Murray), L. interjectum Ching et H. S. Kung ex Ching( = L. japonicum Thunb. ex Murray), L. taliense Ching(＝L. japonicum Thunb. ex Murray), L. pseudoclavatum Ching( = L. japonicm Thunb. ex Murray.), L. pseudoclavatum Ching var. yunnanense Ching( = L. japonicum Thunb. ex Murray), and L. centro-chinense Ching( = L. japonicum Thunb. ex Murray). The distribution of all the taxa is also given. Additionally, some taxonomic discussions are made and it is considered that there is no Diphasiastrum wightianum (Wall. ex Grev. et Hook. ) Holub( = Lycopodium wightianum Wall. ex Grev.et Hook. ) in the flora.     

中国鱼子菜科植物的分类

记述了中国产鱼子菜科(Lemaneaceae)的2属6个种,即鱼子菜属(Lemanea Bory)的中华鱼子菜(L. sinensiaJao)、 粗壮鱼子菜(L. crassa S. L. Xie et Z. X. Shi)、 分枝鱼子菜(L. ramosa S. L. Xie et Z. X. Shi)、 单鱼子菜(L.simplex Jao)和拟鱼子菜属(Paralemanea (Silva) Vis et Sheath)的链状拟鱼子菜(P. catenata (Ktzing) Vis et Sheath)、 小型拟鱼子菜(P. parvula (Sirodot) S. L. Xie et Z. X. Shi)。其中,拟鱼子菜属为中国新记录属,粗壮鱼子菜和分枝鱼子菜为新种,链状拟鱼子菜和小型拟鱼子菜为中国新记录种。     

医药中间体L-核糖的生物制造研究进展

L-核糖(L-ribose)是自然界不存在的L-型稀有糖,L-核糖是多种核苷类似物药物的核心砌块,其衍生物2-脱氧-L-核糖也是昂贵的药物中间体;过去只能通过化学合成获得,近年来开始有酶法合成的报道。本文综述了L-核糖生物制备的三种关键酶-L-阿拉伯糖异构酶(L-AI)、甘露糖-6-磷酸异构酶(MPI)、以及L-核糖异构酶(L-RI)在制备L-核糖方面的研究进展,结合产品衍生物研发进行了总结,并展望了L-核糖在未来医药行业的技术需求。     

Lactobacilli in human dental caries and saliva

Samples (98 plaque and 72 saliva) from 93 patients with dental caries were investigated for Lactobacillus species which comprised 65 (62.5%) of 104 isolates. Yeasts (20.1%), Streptococcus spp. (8.7%), Staphylococcus spp. (2.9%) and a few unidentified species (5.8%), were also found. The Lactobacillus isolates were L. brevis (24.6%) L. fermentum (18.5%) L. casei (16.9%), L. delbrueckii (15.4%), L. plantarum (9.23%), L. acidophilus (7.69%), L. jensenii (4.62%), L. salivarius (1.54%) and L. gasseri (1.54%). The most common species was L. brevis (24.6%). The strains tested for beta-lactamase production showed 75.4% positive. All the Lactobacillus strains were tested for bacteriocin production against Escherichia coli, Salmonella spp., Shigella dysenteriae, S. sonnei, Klebsiella spp. and Campylobacter sp. All the lactobacilli except L. jensenii produced bacteriocin against at least one of the indicator organisms. The involvement of Lactobacillus in dental caries was established, although its role and mechanism is not well understood. The ability of Lactobacillus spp. to protect their host against certain diseases by inhibiting the growth of potential pathogens was evident.     

Paternal inheritance of chloroplast DNA in interspecific hybrids in the genus Larrea (Zygophyllaceae)

The mode of chloroplast DNA (cpDNA) inheritance was investigated in the genus Larrea (Zygophyllaceae) by polymerase chain reaction (PCR) amplification of cpDNA fragments using three pairs of chloroplast universal primers. A total of 20 F(1)s from interspecific crosses among five different taxa in the section Bifolium was examined. Twelve F(1)s were from six crosses between L. cuneifolia (4x) and L. divaricata (2x) (Peru or Argentina) or L. tridentata (2x or 4x). Eight F(1)s were from two sets of reciprocal crosses between L. divaricata (2x) (Argentina) and L. tridentata (2x). Length polymorphism was observed in all three regions of cpDNA that separated L. cuneifolia parents from L. divaricata and L. tridentata parents and in one of the three cpDNA regions that differentiated L. divaricata (Argentina) parents from L. tridentata (2x) parents. In each case, it was the paternal cpDNA marker that appeared in the F(1) individuals. This was further confirmed by restriction fragment length polymorphism (RFLP) analysis of the amplified cpDNA fragments. Larrea may be the fifth genus reported in angiosperms with a paternal bias in cpDNA transmission. Possible mechanisms that may result in paternal cpDNA inheritance were briefly reviewed. Based on the observed uniparental paternal inheritance of cpDNA, restriction analysis of the three cpDNA regions and previous cytogenetic studies, L. divaricata was probably the maternal progenitor of L. cuneifolia.     

中国鱼子菜科植物的分类

记述了中国产鱼子菜科(Lemaneaceae)的2属6个种,即鱼子菜属(Lemanea Bory)的中华鱼子菜(L.sinensiaJao)、粗壮鱼子菜(L.crassa S.L.Xie et Z.X.Shi)、分枝鱼子菜(L.ramosa S.L.Xie et Z.X.Shi)、单鱼子菜(L.simplex Jao)和拟鱼子菜属(Paralemanea(Silva)Vis et Sheath)的链状拟鱼子菜(P.catenata(Kützng)Vis et Sheath)、小型拟鱼子菜(P.parvula(Sirodot)S.L.Xie et Z.X.Shi).其中,拟鱼子菜属为中国新记录属,粗壮鱼子菜和分枝鱼子菜为新种,链状拟鱼子菜和小型拟鱼子菜为中国新记录种.     

Cell population kinetics in pig epidermis: further studies

Abstract. The cell population kinetics of the epidermis were studied in 4-month-old pigs. Mitotic figures were confined to the basal cell (L1) and the first suprabasal cell layer (L2). The mitotic index (MI) was 0.17 ± 0.04% for L1 and 0.08 ± 0.03% for L2. Labelled nuclei were distributed throughout the viable epidermis, the majority (79.1 ± 1.1%) were in L1 with 19.5 ± 1.2% in L2. The labelling indices (LI) in layers L1 and L2 were 7.1 ± 0.4% and 3.4 ± 0.1%, respectively. After labelling with two injections of tritiated thymidine [³H]TdR separated by 90 min, the LI increased to 8.2 ± 0.3% in L1 and to 4.0 ± 0.2% in L2. This increased labelling confirmed that cell proliferation occurs in both layers, L1 and L2, of the epidermis.
The cell production rate ( K ) in L1 and L2 had an upper limit of 10.7 ± 1.0 and 6.2 + 1.8 cells per 1000 cells per hour respectively. The cell flow rate per hour (cell flux), into and out of the DNA synthesis phase (S), and the duration of DNA synthesis were determined from double-labelling studies with [³H]TdR and [¹⁴C]TdR. The cell flux into and out of S was identical and was calculated as 0.6 ± 0.1%/hr (L1) and 0.5 ± 0.1%/hr (L2). Values for t _S varied from 8 to 10 hr. The cell turnover times ( t _T) were in the range 89–129 hr and 180–261 hr for L1 and L2, respectively.
Log normal curves were fitted to the fraction labelled mitoses data for L1 and L2. Values for t _S for cells in L1 and L2 were 9.8 hr and 11.9 hr, respectively. t _G2+ 1/2 t _M was 7.2 hr in L1 and 9.1 hr in L2.